{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:19:47Z","timestamp":1792804787013,"version":"4.5.0"},"reference-count":91,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,28]],"date-time":"2025-12-28T00:00:00Z","timestamp":1766880000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["88887.511314\/2020-00"],"award-info":[{"award-number":["88887.511314\/2020-00"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["407952\/2023-3"],"award-info":[{"award-number":["407952\/2023-3"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Funda\u00e7\u00e3o Amaz\u00f4nia de Amparo a Estudos e Pesquisas","award":["173\/2023"],"award-info":[{"award-number":["173\/2023"]}]},{"name":"Banco da Amaz\u00f4nia","award":["5101 2022\/231"],"award-info":[{"award-number":["5101 2022\/231"]}],"id":[{"id":"https:\/\/ror.org\/05k4nys47","id-type":"ROR","asserted-by":"crossref"}]},{"name":"AQUAGEN\u00d4MICA","award":["443875\/2024-3"],"award-info":[{"award-number":["443875\/2024-3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background\/Objectives: The selection and validation of species-specific housekeeping genes (HKGs) have become increasingly common in functional genomics, with application of quantitative Polymerase Chain Reaction (qPCR) or cDNA-based qPCR (RT-qPCR). Despite the Macrobrachium amazonicum having RNA-seq studies available, there are still no data on the most stable and consistent HKGs for use in relative gene expression analyses. Therefore, the present study aimed to identify and validate seven HKGs in M. amazonicum: Eukaryotic Translation Initiation Factor (EIF), 18S ribosomal RNA (18S), Ribosomal Protein L18 (RPL18), \u03b2-actin, \u03b1-tubulin (\u03b1-tub), Elongation Factor 1-\u03b1 (EF-1\u03b1), and Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH). Methods: The HKGs were identified in the M. amazonicum transcriptome, characterized for identity confirmation, and compared against public databases. Subsequently, RT-qPCR assays were prepared using muscle, hepatopancreas, gills, testis, androgenic gland, and ovary to assess the stability of the HKG markers, employing the comparative \u2206Ct, BestKeeper, NormFinder, and GeNorm methods. Results: All candidate HKGs identified showed high similarity with other decapods. Reactions performed with these markers demonstrated high specificity, PCR efficiency, and elevated coefficients of determination. The comprehensive ranking, indicated that no single HKG was stable across all tissues, with HKGs showing the best stability being tissue-specific. The most stable HKGs were RPL18 and 18S. GAPDH, historically used as an HKG, showed the poorest performance in stability ranking for most tissues tested, whereas \u03b2-actin was most suitable only for ovarian. Conclusions: These data reinforce the need for species-specific HKG validation and provide an appropriate panel of reference markers for gene expression studies in the M. amazonicum.<\/jats:p>","DOI":"10.3390\/genes17010026","type":"journal-article","created":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T00:50:42Z","timestamp":1766969442000},"page":"26","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Identification and Validation of Tissue-Specific Housekeeping Markers for the Amazon River Prawn Macrobrachium amazonicum (Heller, 1862)"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2988-1580","authenticated-orcid":false,"given":"Gabriel Monteiro de","family":"Lima","sequence":"first","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00f4nica Andressa Leite","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00f4mulo Veiga","family":"Paix\u00e3o","sequence":"additional","affiliation":[{"name":"Embrapa Pesca e Aquicultura, Av. NS 10, Cruzamento com a Av. LO 18 Sentido Norte Loteamento-\u00c1gua Fria, Palmas 77008-900, Tocantins, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8664-6440","authenticated-orcid":false,"given":"\u00cdtalo","family":"Lutz","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manoel Alessandro Borges","family":"Aviz","sequence":"additional","affiliation":[{"name":"Instituto Federal de Educa\u00e7\u00e3o, Ci\u00eancia e Tecnologia do Par\u00e1, Campus Camet\u00e1, Av. Euclides Figueiredo, s\/n, Camet\u00e1 68400-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1246-9089","authenticated-orcid":false,"given":"Janieli do Socorro Amorim da Luz","family":"Sousa","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4875-1914","authenticated-orcid":false,"given":"Bruna Ramalho","family":"Maciel","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciano Domingues","family":"Queiroz","sequence":"additional","affiliation":[{"name":"Instituto Federal de Educa\u00e7\u00e3o, Ci\u00eancia e Tecnologia do Par\u00e1, Campus Tucuru\u00ed, Av. Bras\u00edlia, s\/n, Tucuru\u00ed 68455-901, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos Murilo Ten\u00f3rio","family":"Maciel","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iracilda","family":"Sampaio","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0424-2683","authenticated-orcid":false,"given":"Eduardo Sousa","family":"Varela","sequence":"additional","affiliation":[{"name":"Embrapa Pesca e Aquicultura, Av. NS 10, Cruzamento com a Av. LO 18 Sentido Norte Loteamento-\u00c1gua Fria, Palmas 77008-900, Tocantins, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiana Ramalho","family":"Maciel","sequence":"additional","affiliation":[{"name":"Instituto de Estudos Costeiros, Universidade Federal do Par\u00e1, Campus Universit\u00e1rio de Bragan\u00e7a, Al. Leandro Ribeiro, s\/n, Bragan\u00e7a 68600-000, Par\u00e1, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.mam.2005.12.007","article-title":"The Real-time polymerase chain reaction","volume":"27","author":"Kubista","year":"2006","journal-title":"Mol. Asp. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1007\/978-1-4939-3360-0_15","article-title":"Quantitative Real-Time PCR: Recent Advances","volume":"Volume 1392","author":"Luthra","year":"2016","journal-title":"Clinical Applications of PCR"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1677\/jme.0.0250169","article-title":"Absolute quantification of mRNA Using Real-Time Reverse Transcription Polymerase Chain Reaction Assays","volume":"25","author":"Bustin","year":"2000","journal-title":"J. Mol. 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