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1.
Sci Rep ; 14(1): 9455, 2024 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-38658744

RESUMO

The Asian king vulture (AKV), a vital forest scavenger, is facing globally critical endangerment. This study aimed to construct a reference genome to unveil the mechanisms underlying its scavenger abilities and to assess the genetic relatedness of the captive population in Thailand. A reference genome of a female AKV was assembled from sequencing reads obtained from both PacBio long-read and MGI short-read sequencing platforms. Comparative genomics with New World vultures (NWVs) and other birds in the Family Accipitridae revealed unique gene families in AKV associated with retroviral genome integration and feather keratin, contrasting with NWVs' genes related to olfactory reception. Expanded gene families in AKV were linked to inflammatory response, iron regulation and spermatogenesis. Positively selected genes included those associated with anti-apoptosis, immune response and muscle cell development, shedding light on adaptations for carcass consumption and high-altitude soaring. Using restriction site-associated DNA sequencing (RADseq)-based genome-wide single nucleotide polymorphisms (SNPs), genetic relatedness and inbreeding status of five captive AKVs were determined, revealing high genomic inbreeding in two females. In conclusion, the AKV reference genome was established, providing insights into its unique characteristics. Additionally, the potential of RADseq-based genome-wide SNPs for selecting AKV breeders was demonstrated.


Assuntos
Espécies em Perigo de Extinção , Falconiformes , Genoma , Polimorfismo de Nucleotídeo Único , Animais , Falconiformes/genética , Feminino , Variação Genética , Genômica/métodos , Masculino , Tailândia
2.
Evol Appl ; 17(1): e13634, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38283602

RESUMO

Environmental variation can influence the reproductive success of species managed under human care and in the wild, yet the mechanisms underlying this phenomenon remain largely mysterious. Molecular mechanisms such as epigenetic modifiers are important in mediating the timing and progression of reproduction in humans and model organisms, but few studies have linked epigenetic variation to reproductive fitness in wildlife. Here, we investigated epigenetic variation in black-footed ferrets (Mustela nigripes), an endangered North American mammal reliant on ex situ management for survival and persistence in the wild. Despite similar levels of genetic diversity in human-managed and wild-born populations, individuals in ex situ facilities exhibit reproductive problems, such as poor sperm quality. Differences across these settings suggest that an environmentally driven decline in reproductive capacity may be occurring in this species. We examined the role of DNA methylation, one well-studied epigenetic modifier, in this emergent condition. We leveraged blood, testes, and semen samples from male black-footed ferrets bred in ex situ facilities and found tissue-type specificity in DNA methylation across the genome, although 1360 Gene Ontology terms associated with male average litter size shared functions across tissues. We then constructed gene networks of differentially methylated genomic sites associated with three different reproductive phenotypes to explore the putative biological impact of variation in DNA methylation. Sperm gene networks associated with average litter size and sperm count were functionally enriched for candidate genes involved in reproduction, development, and its regulation through transcriptional repression. We propose that DNA methylation plays an important role in regulating these reproductive phenotypes, thereby impacting the fertility of male ex situ individuals. Our results provide information into how DNA methylation may function in the alteration of reproductive pathways and phenotypes in artificial environments. These findings provide early insights to conservation hurdles faced in the protection of this rare species.

3.
Sci Rep ; 13(1): 19806, 2023 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-37957263

RESUMO

Eld's deer, a conserved wildlife species of Thailand, is facing inbreeding depression, particularly in the captive Siamese Eld's deer (SED) subspecies. In this study, we constructed genomes of a male SED and a male Burmese Eld's deer (BED), and used genome-wide single nucleotide polymorphisms to evaluate the genetic purity and the inbreeding status of 35 SED and 49 BED with limited pedigree information. The results show that these subspecies diverged approximately 1.26 million years ago. All SED were found to be purebred. A low proportion of admixed SED genetic material was observed in some BED individuals. Six potential breeders from male SED with no genetic relation to any female SED and three purebred male BED with no relation to more than 10 purebred female BED were identified. This study provides valuable insights about Eld's deer populations and appropriate breeder selection in efforts to repopulate this endangered species while avoiding inbreeding.


Assuntos
Cervos , Polimorfismo de Nucleotídeo Único , Humanos , Animais , Masculino , Feminino , Endogamia , Cervos/genética , Espécies em Perigo de Extinção , Genômica
4.
Sci Rep ; 13(1): 15464, 2023 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-37726331

RESUMO

The critically endangered black rhinoceros (Diceros bicornis; black rhino) experiences extinction threats from poaching in-situ. The ex-situ population, which serves as a genetic reservoir against impending extinction threats, experiences its own threats to survival related to several disease syndromes not typically observed among their wild counterparts. We performed an untargeted metabolomic analysis of serum from 30 ex-situ housed black rhinos (Eastern black rhino, EBR, n = 14 animals; Southern black rhino, SBR, n = 16 animals) and analyzed differences in metabolite profiles between subspecies, sex, and health status (healthy n = 13 vs. diseased n = 14). Of the 636 metabolites detected, several were differentially (fold change > 1.5; p < 0.05) expressed between EBR vs. SBR (40 metabolites), female vs. male (36 metabolites), and healthy vs. diseased (22 metabolites). Results suggest dysregulation of propanoate, amino acid metabolism, and bile acid biosynthesis in the subspecies and sex comparisons. Assessment of healthy versus diseased rhinos indicates involvement of arachidonic acid metabolism, bile acid biosynthesis, and the pentose phosphate pathway in animals exhibiting inflammatory disease syndromes. This study represents the first systematic characterization of the circulating serum metabolome in the black rhinoceros. Findings further implicate mitochondrial and immune dysfunction as key contributors for the diverse disease syndromes reported in ex-situ managed black rhinos.


Assuntos
Doenças do Sistema Imunitário , Metabolômica , Feminino , Masculino , Animais , Síndrome , Perissodáctilos , Ácidos e Sais Biliares
5.
J Hered ; 114(5): 539-548, 2023 08 23.
Artigo em Inglês | MEDLINE | ID: mdl-37249392

RESUMO

The black-footed ferret (Mustela nigripes) narrowly avoided extinction to become an oft-cited example of the benefits of intensive management, research, and collaboration to save a species through ex situ conservation breeding and reintroduction into its former range. However, the species remains at risk due to possible inbreeding, disease susceptibility, and multiple fertility challenges. Here, we report the de novo genome assembly of a male black-footed ferret generated through a combination of linked-read sequencing, optical mapping, and Hi-C proximity ligation. In addition, we report the karyotype for this species, which was used to anchor and assign chromosome numbers to the chromosome-length scaffolds. The draft assembly was ~2.5 Gb in length, with 95.6% of it anchored to 19 chromosome-length scaffolds, corresponding to the 2n = 38 chromosomes revealed by the karyotype. The assembly has contig and scaffold N50 values of 148.8 kbp and 145.4 Mbp, respectively, and is up to 96% complete based on BUSCO analyses. Annotation of the assembly, including evidence from RNA-seq data, identified 21,406 protein-coding genes and a repeat content of 37.35%. Phylogenomic analyses indicated that the black-footed ferret diverged from the European polecat/domestic ferret lineage 1.6 million yr ago. This assembly will enable research on the conservation genomics of black-footed ferrets and thereby aid in the further restoration of this endangered species.


Assuntos
Espécies em Perigo de Extinção , Furões , Animais , Masculino , Furões/genética , Cariótipo , Cariotipagem , Fertilidade
6.
Methods Protoc ; 5(3)2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35736552

RESUMO

Organoids are a type of three-dimensional (3D) cell culture that more closely mimic the in vivo environment and can be maintained in the long term. To date, oviductal organoids have only been reported in laboratory mice, women, and cattle. Equine oviductal organoids were generated and cultured for 42 days (including 3 passages and freeze-thawing at passage 1). Consistent with the reports in mouse and human oviductal organoids, the equine oviductal organoids revealed round cell clusters with a central lumen. Developing a 3D model of the mare oviduct may allow for an increased understanding of their normal physiology, including hormonal regulation. These organoids may provide an environment that mimics the in vivo equine oviduct and facilitate improved in vitro embryo production in equids.

7.
Evol Appl ; 15(3): 351-364, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35386395

RESUMO

Captive breeding programmes represent the most intensive type of ex situ population management for threatened species. One example is the Cuvier's gazelle programme that started in 1975 with only four founding individuals, and after more than four decades of management in captivity, a reintroduction effort was undertaken in Tunisia in 2016, to establish a population in an area historically included within its range. Here, we aim to determine the genetic consequences of this reintroduction event by assessing the genetic diversity of the founder stock as well as of their descendants. We present the first whole-genome sequencing dataset of 30 Cuvier's gazelles including captive-bred animals, animals born in Tunisia after a reintroduction and individuals from a genetically unrelated Moroccan population. Our analyses revealed no difference between the founder and the offspring cohorts in genome-wide heterozygosity and inbreeding levels, and in the amount and length of runs of homozygosity. The captive but unmanaged Moroccan gazelles have the lowest genetic diversity of all genomes analysed. Our findings demonstrate that the Cuvier's gazelle captive breeding programme can serve as source populations for future reintroductions of this species. We believe that this study can serve as a starting point for global applications of genomics to the conservation plan of this species.

8.
Cryobiology ; 104: 56-62, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34788682

RESUMO

Reproductive diseases in mares are a significant cause of subfertility and profound economic loss in the equine industry. Utilizing a 3D in vitro cell culture system that recapitulates the in vivo physiology will reduce time, cost, and welfare concerns associated with in vivo reproductive research in mares. If this 3D model is combined with effective cryopreservation, reproductive research on mares can occur year-round, which is not currently possible in this seasonal species. Endometrial organoids, 3D in vitro cell clusters that exhibit in vivo uterine physiology, have been established in mice, women, and mares. Here we report the first comprehensive assessment of cryopreservation of endometrial organoids in the domestic mare. Organoid growth rate was not affected by the type of freezing media. However, growth rate varied among non-cryopreserved controls, organoids cryopreserved at passage 0 (P0), and organoids cryopreserved at passage 3 (P3). Additionally, there was no difference in organoid viability among freezing media or freezing timepoint (passages). Furthermore, fresh and frozen-thawed organoids displayed positive immunohistochemical staining for ZO-1, which is a marker for intercellular tight junctions, and for periodic acid-Schiff staining as marker for organoid function through mucin production. Results demonstrate that equine endometrial organoids can be cryopreserved with 10% dimethyl sulfoxide with minimal detrimental effects while maintaining intercellular tight junctions (ZO-1) and secretory function. Availability of cryopreserved endometrial organoids may permit expanded research on uterine pathologies that negatively affect mare fertility and improve efficiency, reduce cost, and minimize animal welfare concerns associated with in vivo research in the domestic mare.


Assuntos
Criopreservação , Organoides , Animais , Criopreservação/métodos , Dimetil Sulfóxido/metabolismo , Dimetil Sulfóxido/farmacologia , Endométrio/fisiologia , Feminino , Cavalos , Humanos , Camundongos , Organoides/metabolismo , Útero
9.
J Equine Vet Sci ; 105: 103724, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34607689

RESUMO

The reproductive cycle and early pregnancy represent dynamic physiological states in mammals, but mechanisms involved in early pregnancy in the domestic horse remain poorly understood. Proteins in uterine secretions have been studied, but the proteome of peripheral serum during various reproductive states has not been investigated. This study characterized and compared the serum proteome in the domestic horse during various reproductive states. Serum was collected from three mares during: (1) estrus (day [d] -1; d 0 = ovulation), (2) diestrus (d 12.5, non-mated), (3) early pregnancy (d 12.5, pregnant), and (4) nonpregnant (d 12.5, unsuccessfully mated) states. Serum proteins in each sample were analyzed by Nano LC-MS/MS, and 308 proteins were identified. Differentially-expressed proteins (DEP; > 1.5-fold or < - 0.5-fold) were identified by comparison of protein relative abundance between reproductive states: (1) diestrus compared to estrus (DEP = 71), (2) pregnant compared to diestrus (DEP = 72), and (3) non-pregnant compared to pregnant (DEP = 81). DEPs were analyzed for biological function using PANTHER (pantherdb.org). Several pregnancy-specific proteins previously identified in equine pregnant histotroph, including Apolipoprotein A-I, Complement C3, and Histone H4, were detectable in the serum. The ability to detect these biomarkers in serum provides a more readily available option for investigating and understanding early equine pregnancy.


Assuntos
Proteoma , Espectrometria de Massas em Tandem , Animais , Diestro , Estro , Feminino , Cavalos , Ovulação , Gravidez , Espectrometria de Massas em Tandem/veterinária
10.
Theriogenology ; 172: 106-115, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34153566

RESUMO

The Malayan tapir is a large endangered herbivore native to South-east Asia with fewer than 2500 animals remaining in the wild. Although a small number of animals (183 animals held by 60 institutions) are managed in zoos and breeding centres, there is limited information on the fundamental reproductive biology of this species. The purpose of this present study was to evaluate the associations of reproductive protein biomarkers (CRISP2 and CRISP3) in the seminal plasma and spermatozoa with reproductive characteristics in male Malayan tapirs. Ejaculates were collected from zoo-housed animals by electroejaculation and assessed for sperm motility and quality traits. Seminal plasma and sperm pellets were analysed for CRISP protein expression by immunoblotting. The reproductive tract of a single animal was also analysed for CRISP2 and CRISP3 protein expression and localization by immunohistochemistry. Our results showed that both CRISP2 and CRISP3 are expressed in the seminal plasma and spermatozoa derived from Malayan tapirs. CRISP expression was positively correlated with semen quality, especially ejaculate volume, number of motile sperm, and acrosomal integrity. In addition, CRISP2 and CRISP3 protein expression were slightly high in males that had recently sired an offspring. The results suggest that CRISP proteins may serve as biomarkers for ejaculate quality and fertility in male Malayan tapirs. These findings may have significant implications for planning future breeding and re-introduction efforts for this species.


Assuntos
Análise do Sêmen , Sêmen , Animais , Masculino , Perissodáctilos , Análise do Sêmen/veterinária , Motilidade dos Espermatozoides , Espermatozoides
11.
Anim Reprod Sci ; 226: 106698, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33476905

RESUMO

Large percentages of abnormal sperm, termed teratospermia, are associated with poor fertility in cats, many of which are threatened with extinction from their natural habitats. Even normal appearing spermatozoa from felids with teratospermia may have a compromised capacity for motility and fertilization indicating there are factors affecting the fertilization capacity of all sperm regardless of morphology. There was a comparative study conducted using the RNA-Seq approach to identify differentially expressed genes between morphologically normal and abnormal sperm from domestic cates with normospermia and teratospermia to elucidate genes and pathways associated with abnormal sperm function. Normal sperm from cats with teratospermia have a gene expression profile similar to abnormal sperm from males with teratospermia. There was also downregulation of cGMP pathways which may be associated with a lesser sperm motility in ejaculates from males with teratospermia. Kinase phosphorylation pathways also were downregulated in normal spermatozoa from ejaculates of males with teratospermia. Results indicate that analysis of sperm gene expression provides for a more precise assessment of sperm function in semen of cats with teratospermia and facilitates identification of molecular abnormalities that may lead to compromised fertilization capacity.


Assuntos
Doenças do Gato/genética , Teratozoospermia/veterinária , Animais , Gatos , Criopreservação/veterinária , Regulação da Expressão Gênica , Masculino , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides , Teratozoospermia/genética
12.
Reproduction ; 160(6): 819-831, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33112764

RESUMO

The endometrium, the inner uterine lining, is composed of cell layers that come in direct contact with an embryo during early pregnancy and later with the fetal placenta. The endometrium is responsible for signals associated with normal reproductive cyclicity as well as maintenance of pregnancy. In the mare, functionally competent in vitro models of the endometrium have not been successful. Furthermore, the ability to study various reproductive processes in vitro may allow critical evaluation of signaling pathways involved in the reproductive diseases of animals that cannot be handled frequently, such as various wildlife species. Here we report the establishment of organoids, 3D structures, derived from fresh and frozen-thawed equine endometrium (Equus ferus caballus and E. f. przewalskii). Although organoids from domestic mares responded to exogenous hormonal stimuli, organoids from Przewalski's horse failed to respond to exogenous hormones. The present study represents a 'first' for any large animal model or endangered species. These physiologically functional organoids may facilitate improved understanding of normal reproductive mechanisms, uterine pathologies, and signaling mechanisms between the conceptus and endometrium and may lead to the development of novel bioassays for drug discovery.


Assuntos
Endométrio/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Hormônios/farmacologia , Organoides/efeitos dos fármacos , Animais , Animais Selvagens , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Endométrio/metabolismo , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Feminino , Cavalos , Organoides/metabolismo , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo
13.
Anim Reprod Sci ; 222: 106605, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32987223

RESUMO

The inner layer of the uterus, the endometrium, is responsible and necessary for many reproductive functions. Normal reproductive cyclicity, maternal recognition of pregnancy, maternal interaction with the embryo, and interaction of the reproductive tract with pathogens are dependent on the endometrium. Although most studies have been conducted in vivo using live animals, recent advances in in vitro approaches could facilitate future research in a laboratory setting with minimal effect on animals. Many reproductive studies have been performed in vivo and in vitro in equids, but new in vitro methods to study the endometrium of mares remain unexplored. In this review, there is a description of the normal anatomy and physiology of the mare endometrium in vivo, in vitro endometrial cell culture techniques that have been previously described for the mare, and opportunities for future reproductive research using in vitro methods.


Assuntos
Técnicas de Cultura de Células/veterinária , Endométrio/anatomia & histologia , Endométrio/fisiologia , Cavalos/anatomia & histologia , Cavalos/fisiologia , Animais , Endométrio/citologia , Ciclo Estral/fisiologia , Feminino , Gravidez
14.
Evol Appl ; 13(8): 2143-2154, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32908610

RESUMO

As we enter the sixth mass extinction, many species that are no longer self-sustaining in their natural habitat will require ex situ management. Zoos have finite resources for ex situ management, and there is a need for holistic conservation programs between the public and private sector. Ex situ populations of sable antelope, Hippotragus niger, have existed in zoos and privately owned ranches in North America since the 1910s. Unknown founder representation and relatedness has made the genetic management of this species challenging within zoos, while populations on privately owned ranches are managed independently and retain minimal-to-no pedigree history. Consequences of such challenges include an increased risk of inbreeding and a loss of genetic diversity. Here, we developed and applied a customized targeted sequence capture panel based on 5,000 genomewide single-nucleotide polymorphisms to investigate the genomic diversity present in these uniquely managed populations. We genotyped 111 sable antelope: 23 from zoos, 43 from a single conservation center, and 45 from ranches. We found significantly higher genetic diversity and significantly lower inbreeding in herds housed in zoos and conservation centers, when compared to those in privately owned ranches, likely due to genetic-based breeding recommendations implemented in the former populations. Genetic clustering was strong among all three populations, possibly as a result of genetic drift. We propose that the North American ex situ population of sable antelope would benefit from a metapopulation management system, to halt genetic drift, reduce the occurrence of inbreeding, and enable sustainable population sizes to be managed ex situ.

15.
Case Rep Vet Med ; 2020: 8810770, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32724700

RESUMO

Dama gazelles (Nanger dama mhorr) are considered critically endangered by the IUCN Red List of Threatened Species since the wild population is comprised of fewer than 200 animals. Several zoos in Europe and some private ranches in the US (Texas) maintain this species in captivity as a hedge against extinction. A routine reproductive exam on an adult male revealed bilateral cysts in the testicular parenchyma. The fluid in the cysts contained copious, moderately progressive motile spermatozoa. Following examination, the gazelle was diagnosed with bilateral intratesticular spermatoceles. Electroejaculation yielded 1.5 ml of semen containing 387 × 106 total sperm with 50% motility and 45% morphologically normal sperm. The spermatoceles did not appear to cause pain or dysfunction, so no treatment was performed at this time. Since fewer than 10 animals are managed in captivity in the US, no intervention (castration/hemicastration) was considered.

16.
Anim Reprod Sci ; 217: 106459, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32408971

RESUMO

Assisted reproduction of endangered equids, such as Persian onagers (Equus hemionus onager), is vital for species conservation. Little is known about Persian onager reproductive functions, including functions of the uterine endometrium. Recently, successful cryopreservation of the domestic mare endometrium was reported, but there is no information on cryo-sensitivity or in vitro culture of endometrial tissues of any non-domestic equid. In the present study, endometrial explants from Persian onagers were cryopreserved and cultured in vitro for 5 days. There was no difference between endometrial explants when 10% and 20% dimethyl sulfoxide (DMSO) was used for cryopreservation. Cell viability and structural integrity were comparable to fresh tissue. Abundance of estrogen receptor-α (ESR1) and progesterone receptor (PGR) mRNA transcript in endometrial explants was less in most treatment groups compared to the fresh tissue control. There was variation in E-cadherin mRNA abundance in endometrial explants among treatment groups with some treatment groups having a lesser abundance compared to the control group. The abundance of Ki67 mRNA transcript of endometrial explants was not different among treatment groups compared to the control group. Results indicate that DMSO is a suitable cryoprotectant for the Persian onager endometrium, and in vitro culture in a liquid-gas interface can maintain Persian onager endometrial explants for as long as 5 days. Findings allow for a greater understanding of reproductive mechanisms in vitro for this endangered species and other domestic equids including donkeys.


Assuntos
Criopreservação/veterinária , Endométrio , Equidae/fisiologia , Técnicas de Cultura de Tecidos/veterinária , Preservação de Tecido/veterinária , Animais , Feminino
17.
J Equine Vet Sci ; 88: 102938, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32303306

RESUMO

Insulin resistance (IR) is characterized by an increase in biomarkers of systemic inflammation and susceptibility to laminitis in horses. Impacts on reproduction include a lengthened interovulatory period in horses. Dietary omega-3 (docosahexaenoic acid [DHA]) promotes anti-inflammatory processes, has been implicated in health benefits, and can reduce cytokine secretion. This preliminary study investigated the impact of IR as well as the influence of dietary supplementation (DHA) on the uterine fluid proteome in early pregnant horses. Mares were artificially inseminated; uterine fluid and embryos were collected on d 12.5 after ovulation. Uterine fluid was pooled for metabolic and diet categories (n = 8; n = 2 per metabolic and dietary status) and concentrated, and the proteome was analyzed using tandem mass spectrometry (iTRAQ). Five proteins met differential abundance criteria (±1.5-fold change, P < .05) in all comparisons (Control C, IS vs. C, IR; C, IS vs. DHA, IS; C, IR vs. DHA, IR). Serum amyloid A, afamin, and serotransferrin were upregulated in C, IR mares but downregulated in DHA, IR mares when compared to C, IS and C, IR, respectively. Quantitative PCR supported mass spectrometry results. The presence of serum amyloid A and serotransferrin in histotroph of IR mares potentially indicates an inflammatory response not seen in IS counterparts. These preliminary findings provide novel evidence on the potential impact of insulin resistance and DHA supplementation on the secreted equine uterine proteome during early pregnancy.


Assuntos
Ácidos Graxos Ômega-3 , Proteoma , Animais , Dieta/veterinária , Ácidos Docosa-Hexaenoicos , Feminino , Cavalos , Gravidez , Espectrometria de Massas em Tandem/veterinária
18.
Cryobiology ; 91: 90-96, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31626783

RESUMO

Availability of viable frozen-thawed endometrial tissues could facilitate detailed studies into physiologic and disease processes influencing the endometrium. This study was designed to investigate the cryosurvival of equine endometrial tissue. Previous studies in the human and horse have focused on cryopreservation of dissociated endometrial cells. To our knowledge, there are no studies on cryopreservation of endometrial explants. Our objectives were to 1) determine the influence of differing concentrations of the permeating cryoprotectant dimethyl sulfoxide (Me2SO) on viability, structural integrity, and gene expression of cryopreserved equine endometrial tissues prior to and following a 5-day explant culture in vitro and 2) examine the influence of low (1000 mg/L dextrose) vs high (4500 mg/L dextrose) glucose medium during in vitro culture. Both 10% and 20% (v/v) concentrations of Me2SO maintained viability following cryopreservation and in vitro culture. In addition, gene expression remained unaltered following cryopreservation with either 10% or 20% Me2SO. However, tissue structural integrity was slightly reduced compared to the fresh control. Furthermore, there was no difference in structural integrity, cell viability, or gene expression between low and high glucose medium during in vitro culture. Although E-cadherin and Ki67 gene expression was not different among fresh, 10% Me2SO, and 20% Me2SO treatments prior to or following tissue culture, estrogen receptor-α and progesterone receptor gene expression were reduced in all groups after explant culture. This is the first report of successful cryopreservation of equine endometrial explants.


Assuntos
Criopreservação/veterinária , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Endométrio/fisiologia , Cavalos/fisiologia , Animais , Sobrevivência Celular , Criopreservação/métodos , Feminino , Humanos
19.
PLoS One ; 14(7): e0217986, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31269032

RESUMO

There is limited physiological information on onset of puberty in male lions. The aim of this study was to use longitudinal non-invasive monitoring to: 1) assess changes in steroid metabolite excretory patterns as a function of age and body weight; 2) determine correlations between fecal androgen (FAM) and glucocorticoid (FGM) metabolite concentrations; and 3) confirm spermiogenesis non-invasively through urinalysis. Specifically, FAM and FGM metabolites were analyzed in samples collected twice weekly from 21 male lions at 17 institutions (0.9-16 years of age) for 3.8 months- 2.5 years to assess longitudinal hormone patterns. In addition, body weights were obtained approximately monthly from 10 individuals at five zoos (0.0-3.0 years), and urine was collected from six males at two facilities (1.2-6.3 years) and evaluated for the presence of spermatozoa. An increase in overall mean FAM occurred at 2.0 years of age, at which point concentrations remained similar throughout adulthood. The onset of puberty occurred earlier in captive-born males (<1.2 years of age) compared to wild-born counterparts (<2.5 years of age). Additionally, males in captivity gained an average of 7.3 kg/month compared to 3.9 kg/month for wild males over the first 2-2.5 years of age. Sperm (spermaturia) was observed in males as young as 1.2 years in captivity compared to 2.5 years in the wild (ejaculates). There was no difference in FAM or FGM concentrations with regards to age or season. Overall, this study demonstrates that: 1) captive male lions attain puberty at an earlier age than wild counterparts; 2) onset of puberty is influenced by body weight (growth rate); and 3) spermiogenesis can be confirmed via urinalysis. Knowledge about the linkage between body weight and onset of puberty could facilitate improved reproductive management of ex situ populations via mitigating the risk of unintended breedings in young animals.


Assuntos
Androgênios/metabolismo , Peso Corporal/fisiologia , Glucocorticoides/metabolismo , Leões/fisiologia , Maturidade Sexual/fisiologia , Espermatozoides/metabolismo , Animais , Fezes , Masculino , Espermatozoides/citologia
20.
G3 (Bethesda) ; 9(6): 1785-1793, 2019 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-31000506

RESUMO

Genome-wide assessment of genetic diversity has the potential to increase the ability to understand admixture, inbreeding, kinship and erosion of genetic diversity affecting both captive (ex situ) and wild (in situ) populations of threatened species. The sable antelope (Hippotragus niger), native to the savannah woodlands of sub-Saharan Africa, is a species that is being managed ex situ in both public (zoo) and private (ranch) collections in the United States. Our objective was to develop whole genome sequence resources that will serve as a foundation for characterizing the genetic status of ex situ populations of sable antelope relative to populations in the wild. Here we report the draft genome assembly of a male sable antelope, a member of the subfamily Hippotraginae (Bovidae, Cetartiodactyla, Mammalia). The 2.596 Gb draft genome consists of 136,528 contigs with an N50 of 45.5 Kbp and 16,927 scaffolds with an N50 of 4.59 Mbp. De novo annotation identified 18,828 protein-coding genes and repetitive sequences encompassing 46.97% of the genome. The discovery of single nucleotide variants (SNVs) was assisted by the re-sequencing of seven additional captive and wild individuals, representing two different subspecies, leading to the identification of 1,987,710 bi-allelic SNVs. Assembly of the mitochondrial genomes revealed that each individual was defined by a unique haplotype and these data were used to infer the mitochondrial gene tree relative to other hippotragine species. The sable antelope genome constitutes a valuable resource for assessing genome-wide diversity and evolutionary potential, thereby facilitating long-term conservation of this charismatic species.


Assuntos
Antílopes/genética , Genoma , Genômica , Sequenciamento Completo do Genoma , Animais , Antílopes/classificação , Biodiversidade , Evolução Biológica , Biologia Computacional/métodos , Feminino , Variação Genética , Genética Populacional , Genoma Mitocondrial , Genômica/métodos , Masculino , Anotação de Sequência Molecular , Fenótipo , Filogenia , Estados Unidos
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