{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T08:52:17Z","timestamp":1765961537108,"version":"build-2065373602"},"reference-count":60,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,5,13]],"date-time":"2022-05-13T00:00:00Z","timestamp":1652400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Center, Poland","doi-asserted-by":"publisher","award":["2014\/15\/B\/NZ5\/O3572","010\/RID\/2018\/19"],"award-info":[{"award-number":["2014\/15\/B\/NZ5\/O3572","010\/RID\/2018\/19"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004569","name":"Minister of Education and Science","doi-asserted-by":"publisher","award":["2014\/15\/B\/NZ5\/O3572","010\/RID\/2018\/19"],"award-info":[{"award-number":["2014\/15\/B\/NZ5\/O3572","010\/RID\/2018\/19"]}],"id":[{"id":"10.13039\/501100004569","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Uterine inflammation is a common pathology in animals, leading to disturbances in reproductive processes and reduced production profitability. Pituitary adenylate cyclase-activating peptide (PACAP) effects at the uterine level during inflammation are not known. In the current study, we analyzed the relative PACAP type 1 receptor (PAC1R) mRNA transcript and protein abundances in the myometrium (MYO), as well s PACAP and PAC1R involvement in the contractile function of inflamed pig uterus. To that end, E. coli suspension (E. coli group) or saline (SAL group) was injected into the uterine horns or laparotomy was performed (CON group). Eight days after the bacteria injections, severe acute endometritis and a reduced relative abundance of PAC1R protein in the MYO were observed. Compared to the period before PACAP in vitro administration, PACAP (10\u22127 M) in the CON and SAL groups decreased in amplitude in the MYO and endometrium (ENDO)\/MYO, whereas in the E. coli group, increased amplitude in the MYO and reduced amplitude in the ENDO\/MYO were observed. In the E. coli group, PACAP enhanced the amplitude in the MYO (10\u22127 M) and decreased the amplitude in the ENDO\/MYO (10\u22128 M) compared with other groups. PACAP (10\u22127 M) increased the frequency of both kinds of strips in the CON and SAL groups compared with the pretreatment period. PACAP (both doses) did not significantly change the frequency in the E. coli group, whereas in response to PACAP (10\u22127 M), the frequency was reduced compared to other groups. In the MYO, PAC1R antagonist decreased the amplitude reduction (CON and SAL groups) and reversed a rise in PACAP (10\u22127 M)-evoked amplitude (E. coli group). PAC1R blocking reversed (MYO) and abolished (ENDO\/MYO) the stimulatory effect of PACAP (10\u22127 M) on the frequency (CON and SAL groups). PAC1R antagonist and PACAP (10\u22127 M) evoked the appearance of frequency depression in both kinds of strips (E. coli group). In summary, in pigs, severe acute endometritis reduces the relative abundance of PAC1R protein in the MYO, and PAC1R mediates the influence of PACAP on inflamed uterus contractility.<\/jats:p>","DOI":"10.3390\/ijms23105467","type":"journal-article","created":{"date-parts":[[2022,5,14]],"date-time":"2022-05-14T09:27:56Z","timestamp":1652520476000},"page":"5467","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Investigation of the Role of Pituitary Adenylate Cyclase-Activating Peptide (PACAP) and Its Type 1 (PAC1) Receptor in Uterine Contractility during Endometritis in Pigs"],"prefix":"10.3390","volume":"23","author":[{"given":"Barbara","family":"Jana","sequence":"first","affiliation":[{"name":"Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima 10, 10-078 Olsztyn, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaros\u0142aw","family":"Ca\u0142ka","sequence":"additional","affiliation":[{"name":"Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury, Oczapowskiego 13, 10-718 Olsztyn, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Krzysztof","family":"Witek","sequence":"additional","affiliation":[{"name":"Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima 10, 10-078 Olsztyn, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1095\/biolreprod.109.077370","article-title":"Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle","volume":"81","author":"Sheldon","year":"2009","journal-title":"Biol. Reprod."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1007\/s00580-009-0929-1","article-title":"Endometritis in gilts: Reproductive data, bacterial culture, histopathology, and infiltration of immune cells in the endometrium","volume":"19","author":"Tummaruk","year":"2010","journal-title":"Comp. Clin. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13028-015-0175-2","article-title":"Periparturient stress and immune suppression as a potential cause of retained placenta in highly productive dairy cows: Examples of prevention","volume":"57","author":"Mordak","year":"2015","journal-title":"Acta Vet. Scand."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.theriogenology.2020.01.042","article-title":"Modulation of immune function in the bovine uterus peripartum","volume":"150","author":"Pascottini","year":"2020","journal-title":"Theriogenology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1111\/rda.13862","article-title":"In vitro effects of lipopolysaccharides on bovine uterine contractility","volume":"56","author":"Wiebe","year":"2021","journal-title":"Reprod. Domest. Anim."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1016\/j.theriogenology.2015.12.022","article-title":"The effect of puerperal uterine disease on histopathologic findings and mRNA expression of proinflammatory cytokines of the endometrium in dairy cows","volume":"85","author":"Heppelmann","year":"2016","journal-title":"Theriogenology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.jri.2016.07.002","article-title":"Innate immunity and the sensing of infection, damage and danger in the female genital tract","volume":"119","author":"Sheldon","year":"2017","journal-title":"J. Reprod. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"654","DOI":"10.22358\/jafs\/66822\/2007","article-title":"The influence of inflammatory process on prostaglandin F2\u03b1 contractile activity in porcine uterus","volume":"16","author":"Kucharski","year":"2007","journal-title":"J. Anim. Feed Sci."},{"key":"ref_9","first-page":"335","article-title":"Participation of prostaglandin E2 in contractile activity of inflamed porcine uterus","volume":"79","author":"Jana","year":"2010","journal-title":"Acta Vet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1016\/j.theriogenology.2012.10.020","article-title":"Synthesis of prostacyclin and its effect on the contractile activity of the inflamed porcine uterus","volume":"79","author":"Jana","year":"2013","journal-title":"Theriogenology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1328","DOI":"10.1016\/j.theriogenology.2015.01.021","article-title":"The influence of leukotrienes C4 and D4 on the contractility of an inflamed porcine uterus","volume":"83","author":"Jana","year":"2015","journal-title":"Theriogenology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.anireprosci.2017.11.019","article-title":"Effects of oxytocin and PGF2\u03b1 on uterine contractility in cows with and without metritis-An in-vitro study","volume":"188","author":"Heppelmann","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.theriogenology.2018.06.025","article-title":"Expression of alpha and beta adrenergic receptors in the pig uterus during inflammation","volume":"119","author":"Meller","year":"2018","journal-title":"Theriogenology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"11512","DOI":"10.1038\/s41598-021-91184-5","article-title":"Role of beta-adrenergic receptor subtypes in pig uterus contractility with inflammation","volume":"11","author":"Jana","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.theriogenology.2022.01.025","article-title":"Roles of alpha-2-adrenergic receptor isoforms in inflamed pig uterus contractility in vitro","volume":"183","author":"Jana","year":"2022","journal-title":"Theriogenology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.theriogenology.2019.09.015","article-title":"Participation of acetylcholine and its receptors in the contractility of inflamed porcine uterus","volume":"143","author":"Jana","year":"2020","journal-title":"Theriogenology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"531","DOI":"10.2478\/jvetres-2020-0073","article-title":"Escherichia coli-induced inflammation changes the expression of acetylcholine receptors (M2R, M3R, and \u03b1-7 nAChR) in pig uterus","volume":"64","author":"Jana","year":"2020","journal-title":"J. Vet. Res."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Jana, B., Ca\u0142ka, J., and Palus, K. (2020). Inflammation changes the expression of neuropeptide Y receptors in the pig myometrium and their role in the uterine contractility. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0236044"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.rvsc.2020.09.016","article-title":"The role of somatostatin and its receptors (sstr2, sstr5) in the contractility of gilt inflamed uterus","volume":"133","author":"Jana","year":"2020","journal-title":"Res. Vet. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"106680","DOI":"10.1016\/j.anireprosci.2020.106680","article-title":"Alterations in the relative abundance of the vasoactive intestinal peptide receptors (VPAC1 and VPAC2) and functions in uterine contractility during inflammation","volume":"225","author":"Palus","year":"2021","journal-title":"Anim. Reprod. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Jana, B., Ca\u0142ka, J., and Mici\u0144ski, B. (2021). Regulatory influence of galanin and GALR1\/GALR2 receptors on inflamed uterus contractility in pigs. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22126415"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"644644","DOI":"10.3389\/fmolb.2021.644644","article-title":"Molecular Basis of Class B GPCR Selectivity for the Neuropeptides PACAP and VIP","volume":"8","author":"Liao","year":"2021","journal-title":"Front. Mol. Biosci."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Karpiesiuk, A., and Palus, K. (2021). Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) in Physiological and Pathological Processes within the Gastrointestinal Tract: A Review. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22168682"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1097\/MED.0000000000000621","article-title":"Pituitary adenylate cyclase-activating polypeptide\/vasoactive intestinal peptide (Part 2): Biology and clinical importance in central nervous system and inflammatory disorders","volume":"28","author":"Moody","year":"2021","journal-title":"Curr. Opin. Endocrinol. Diabetes Obes."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.rvsc.2019.05.003","article-title":"Endometritis affects chemical coding of the dorsal root ganglia neurons supplying uterus in the sexually mature gilts","volume":"124","author":"Bulc","year":"2019","journal-title":"Res. Vet. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/j.peptides.2005.08.005","article-title":"Expression of pituitary adenylate cyclase activating peptide in the uterine cervix, lumbosacral dorsal root ganglia and spinal cord of rats during pregnancy","volume":"27","author":"Papka","year":"2006","journal-title":"Peptides"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Podlasz, P., and Wasowicz, K. (2021). Effect of partial hysterectomy on the neurons of the paracervical ganglion (PCG) of the pig. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0253676"},{"key":"ref_28","first-page":"E108","article-title":"Pituitary adenylate cyclase-activating polypeptide: Occurrence and relaxant effect in female genital tract","volume":"269","author":"Steenstrup","year":"1995","journal-title":"Am. J. Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1016\/0306-4522(96)00082-6","article-title":"Pituitary adenylate cyclase activating polypeptide innervation of the rat female reproductive tract and the associated paracervical ganglia: Effect of capsaicin","volume":"73","author":"Fahrenkrug","year":"1996","journal-title":"Neuroscience"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Rytel, L., and Gonkowski, S. (2020). The influence of bisphenol A on the nitrergic nervous structures in the domestic porcine uterus. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21124543"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1111\/j.1748-1716.1994.tb09792.x","article-title":"Pituitary adenylate cyclase activating polypeptide induces vascular relaxation and inhibits non-vascular smooth muscle activity in the rabbit female genital tract","volume":"152","author":"Steenstrup","year":"1994","journal-title":"Acta Physiol. Scand."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1007\/s12031-014-0337-0","article-title":"Investigation of the possible functions of PACAP in human trophoblast cells","volume":"54","author":"Horvath","year":"2014","journal-title":"J. Mol. Neurosci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.repbio.2020.01.005","article-title":"Possible effects of pituitary adenylate cyclase activating polypeptide (PACAP) on early embryo implantation marker HB-EGF in mouse","volume":"20","author":"Cseh","year":"2020","journal-title":"Reprod. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1002\/pmic.201000320","article-title":"Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies","volume":"11","author":"Verma","year":"2011","journal-title":"Proteomics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1677\/joe.1.06470","article-title":"Characterization and expression of different pituitary adenylate cyclase-activating polypeptide\/vasoactive intestinal polypeptide receptors in rat ovarian follicles","volume":"191","author":"Vaccari","year":"2006","journal-title":"J. Endocrinol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1453","DOI":"10.1016\/S0196-9781(97)00142-3","article-title":"Rat corpus luteum expresses both PACAP and PACAP type IA receptor mRNAs","volume":"18","author":"Kotani","year":"1997","journal-title":"Peptides"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1158","DOI":"10.1210\/endo.141.3.7346","article-title":"Pituitary adenylate cyclase-activating polypeptide (PACAP) and PACAP-receptor type 1 expression in rat and human placenta","volume":"141","author":"Scaldaferri","year":"2000","journal-title":"Endocrinology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1002\/mrd.10221","article-title":"Expression patterns of pituitary adenylate cyclase activating polypeptide and its type I receptor mRNAs in the rat placenta","volume":"64","author":"Koh","year":"2003","journal-title":"Mol. Reprod. Dev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1007\/s12031-008-9104-4","article-title":"PACAP-mediated ATP release from rat urothelium and regulation of PACAP\/VIP and receptor mRNA in micturition pathways after cyclophosphamide (CYP)-induced cystitis","volume":"36","author":"Girard","year":"2008","journal-title":"J. Mol. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1111\/j.1365-2222.2010.03636.x","article-title":"Pituitary adenylate cyclase-activating peptide receptor 1 mediates anti-inflammatory effects in allergic airway inflammation in mice","volume":"41","author":"Lauenstein","year":"2011","journal-title":"Clin. Exp. Allergy"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/0167-0115(95)00156-5","article-title":"Pituitary adenylate cyclase activating polypeptide (PACAP): Occurrence and vasodilatory effect in the human uteroplacental unit","volume":"61","author":"Steenstrup","year":"1996","journal-title":"Regul. Pept."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10097","DOI":"10.1002\/jcb.28293","article-title":"PACAP through EGFR transactivation preserves human corneal endothelial integrity","volume":"120","author":"Maugeri","year":"2019","journal-title":"J. Cell. Biochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3245","DOI":"10.1161\/01.CIR.0000147235.53360.59","article-title":"Pulmonary hypertension and right heart failure in pituitary adenylate cyclase-activating polypeptide type I receptor-deficient mice","volume":"110","author":"Otto","year":"2004","journal-title":"Circulation"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.nurt.2010.02.003","article-title":"The PACAP receptor: A novel target for migraine treatment","volume":"7","author":"Schytz","year":"2010","journal-title":"Neurotherapeutics"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/0196-9781(93)90149-B","article-title":"Pituitary adenylate cyclase activating polypeptide (PACAP) is a potent relaxant of the rat ileum","volume":"14","author":"Katsoulis","year":"1993","journal-title":"Peptides"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1111\/j.1749-6632.1996.tb17497.x","article-title":"Role of PACAP in the regulation of gastrointestinal motility","volume":"805","author":"Katsoulis","year":"1996","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1111\/j.1476-5381.1996.tb15719.x","article-title":"VIP- and PACAP-mediated nonadrenergic, noncholinergic inhibition in longitudinal muscle of rat distal colon: Involvement of activation of charybdotoxin- and apamin-sensitive K+ channels","volume":"119","author":"Kishi","year":"1996","journal-title":"Br. J. Pharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/S0014-2999(97)01144-8","article-title":"Distinct receptors mediate pituitary adenylate cyclase-activating peptide- and vasoactive intestinal peptide-induced relaxation of rat ileal longitudinal muscle","volume":"334","author":"Ekblad","year":"1997","journal-title":"Eur. J. Pharmacol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1038\/sj.bjp.0705582","article-title":"Heterogeneity of neuronal and smooth muscle receptors involved in the VIP- and PACAP-induced relaxations of the pig intravesical ureter","volume":"141","author":"Barahona","year":"2004","journal-title":"Br. J. Pharmacol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"R951","DOI":"10.1152\/ajpregu.00734.2005","article-title":"Role for pituitary adenylate cyclase activating polypeptide in cystitis-induced plasticity of micturition reflexes","volume":"290","author":"Braas","year":"2006","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1677\/joe.0.1520141","article-title":"Presence of pituitary adenylate cyclase-activating polypeptide 38-immuno-like material in the brain and ovary of the female crested newt, Triturus carnifex: Its involvement in the ovarian synthesis of prostaglandins and steroids","volume":"152","author":"Gobbetti","year":"1997","journal-title":"J. Endocrinol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1161\/01.HYP.34.5.1152","article-title":"Time-dependent effects of the neuropeptide PACAP on catecholamine secretion: Stimulation and desensitization","volume":"34","author":"Taupenot","year":"1999","journal-title":"Hypertension"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.mcn.2009.11.007","article-title":"PACAP\/PAC1R signaling modulates acetylcholine release at neuronal nicotinic synapses","volume":"43","author":"Pugh","year":"2010","journal-title":"Mol. Cell. Neurosci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.neuropharm.2003.09.011","article-title":"Interplay between PACAP and NO in mouse ileum","volume":"46","author":"Zizzo","year":"2004","journal-title":"Neuropharmacology"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.anireprosci.2006.01.002","article-title":"Changes in prostaglandin production and ovarian function in gilts during endometritis induced by Escherichia coli infection","volume":"97","author":"Jana","year":"2007","journal-title":"Anim. Reprod. Sci."},{"key":"ref_56","first-page":"207","article-title":"Nitric oxide mediates an inflammatory effect of Escherichia coli in the porcine uterus","volume":"4","author":"Jana","year":"2000","journal-title":"Pol. J. Vet. Sci."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1089\/cmb.2005.12.1047","article-title":"Comprehensive algorithm for quantitative real-time polymerase chain reaction","volume":"12","author":"Zhao","year":"2005","journal-title":"J. Comput. Biol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","article-title":"Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets","volume":"64","author":"Andersen","year":"2004","journal-title":"Cancer Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1007\/BF00318601","article-title":"The origin of ovarian neuropeptide Y (NPY)-immunoreactive nerve fibres from the inferior mesenteric ganglion in the pig","volume":"266","author":"Majewski","year":"1991","journal-title":"Cell Tissue Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/10\/5467\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:10:32Z","timestamp":1760137832000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/10\/5467"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,13]]},"references-count":60,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,5]]}},"alternative-id":["ijms23105467"],"URL":"https:\/\/doi.org\/10.3390\/ijms23105467","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2022,5,13]]}}}