EFEITO DO CONTROLE DE LUZ SOBRE AS CARACTERÍSTICAS ESPERMÁTICAS E PROTEÔMICA DO SÊMEN DE PERDIZES (Rhynchotus rufescens)

RESUMO

O estudo teve como objetivo avaliar as características seminais e o comprimento da estação reprodutiva de perdizes (Rhynchotus rufescens), criadas com luz natural ou artificial, a fim de melhorar a eficiência reprodutiva visando à criação comercial (estudo I). Foi também objetivo descrever a proteômica dos espermatozoides e plasma seminal de perdizes (estudo II). No estudo I, durante a estação reprodutiva (setembro 2017 a março 2018), foram selecionados 20 de 60 machos, quanto à facilidade para a colheita do sêmen e à produção espermática. Os machos selecionados foram divididos em 2 grupos, grupo controle (luz natural, n = 10) e grupo tratado (16 h de luz e 8 h de escuridão, n = 10). O sêmen foi colhido 2 vezes/semana e analisado quanto ao volume, motilidade subjetiva, vigor, coloração, grau de contaminação, concentração espermática e defeitos morfológicos. O estudo II foi conduzido similarmente ao estudo I. O sêmen foi colhido de 28 machos, divididos em 2 grupos (luz natural e luz artificial). Após as colheitas, as amostras foram agrupadas em pools de cada grupo e centrifugadas para a separação do plasma seminal (sobrenadante) e espermatozoides (pellet). As amostras foram preparadas para proteômica com digestão tríptica e submetidas a espectrometria de massas (ESI-Q-TOF MS/MS). No estudo I, a obtenção e facilidade na colheita do sêmen foi maior nas aves mantidas com luz artificial (43 versus 11 amostras de sêmen dos grupos luz artificial e natural, respectivamente). Os parâmetros avaliados no sêmen, também foram superiores (P < 0,001) nas aves mantidas sob luz artificial. No estudo II, foram encontradas 28 proteínas no plasma seminal e 58 no extrato proteico dos espermatozoides. A keratin e isoformas e as catalytic activity e binding foi a proteína e funções moleculares mais frequentes, respectivamente, nas amostras e grupos. No extrato proteico dos espermatozoides, as proteínas alpha-enolase, serum albumin, keratin, type II cytoskeletal cochleal (fragment) e ferritin (fragment) foram encontradas em maior abundância nas aves mantidas com luz natural. Já as proteínas olfactory receptor (fragment), tail-anchored protein insertion receptor WRB (fragment), keratin, type II cytoskeletal 73 (fragment) e keratin, type II cytoskeletal 75 (fragment) foram mais abundantes nas aves com luz artificial. As proteínas identificadas no plasma seminal foram keratin, type II cytoskeletal 75 frag., keratin, type I cytoskeletal 15, keratin, type I cytoskeletal 17, keratin, type II cytoskeletal 73, keratin, type II cytoskeletal 75, keratin, type II cytoskeletal cochleal, tubulin beta chain (Fragment), glutathione S-transferase Mu 5. Nós concluímos que o programa de luz artificial pode prolongar a estação reprodutiva e aumentar a qualidade espermática de perdizes, sendo indicado para a produção comercial. Além disso, as principais proteínas das células espermáticas e plasma seminal de perdizes (Rhynchotus rufescens) foram descritas pela primeira vez, as quais foram similares as encontradas em outras espécies.

ABSTRACT

The study aimed to evaluate the seminal characteristics and reproductive breeding length of the red-winged tinamou (Rhynchotus rufescens), breed with natural and artificial light, in order to improve reproductive efficiency to commercial breeding (study I). Also, it was aim to describe the sperm and seminal plasma proteomic of tinamou (estudy II). In the suty I, during breeding season (Setember 2017 to March 2018), 20 males were select from 60 males, regarding the ease of semen collection and sperm production. The selected males were divided into 2 groups, control group (natural light, n = 10) and treated group (16 h light and 8 h darkness, n = 10). Semen was collected two time/week and analyzed for volume, subjective motility, vigor, color, contamination degree, sperm concentration and the morphological defects. The study II was similarly conducted to study I. The semen was collected from 28 males, divided in 2 groups (artificial and natural light). After the collections, the samples was grouped in pools of the each groups and centrifuged to separate seminal plasma (supernatant) and spermatozoa (pellet). The samples were prepared to proteomic with triptic digestion and submitted to mass spectrometry. In the study I, there was higher samples number and easiness to collect the sample during in the birds maintained to the artificial light (43 versus 11 from artificial and natural light groups, respectively). The parameters evaluated in semen were also higher (P < 0.001) in birds kept under artificial light. In the study II, 28 proteins were found in the seminal plasma and 58 in the protein extract of the spermatozoa. Keratin and isoforms and catalytic activity and binding were the most frequent proteins and molecular functions, respectively, in samples and groups. In the protein extract of sperm, the proteins alpha-enolase, serum albumin, keratin, type II cytoskeletal cochleal (fragment) and ferritin (fragment) were found in greater abundance in birds kept under natural light. The olfactory receptor proteins (fragment), tail-anchored protein insertion receptor WRB (fragment), keratin, type II cytoskeletal 73 (fragment) and keratin, type II cytoskeletal 75 (fragment) were more abundant in birds with artificial light. The main proteins identified in the seminal plasma were keratin, type II cytoskeletal 75 frag., Keratin, type I cytoskeletal 15, keratin, type I cytoskeletal 17, keratin, type II cytoskeletal 73, keratin, type II cytoskeletal 75, keratin, type II cytoskeletal cochleal , tubulin beta chain (Fragment), glutathione S-transferase Mu 5. In conclusion, the artificial light program can prolong the reproductive season and increase the sperm quality of red-winged tinamou, being indicated for commercial production. Moreover, the main proteins of sperm cells and seminal plasma of red-winged tinamou (Rhynchotus rufescens) were described for first time and were found similar proteins to other species.