Giant panda (<i>Ailuropoda melanoleuca</i>) sperm morphometry and function after repeated freezing and thawing

Santiago-Moreno, J.; Esteso, M.C.; Pradieé, J.; Castaño, C.; Toledano-Dı́az, A.; O’Brien, E.; López-Sebastián, A.; Martínez‐Nevado, Eva; Delclaux, M.; Fernández-Morán, J.; Zhihe, Zheng

doi:10.1111/and.12468

Cited by 26 publications

(19 citation statements)

References 17 publications

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“…It has also been suggested that depolarization of sperm membrane associated with cAMP maybe an important change in sperm membrane during cryopreservation [41]. Compared to boar sperm, giant panda sperm appears to be strongly cryo-tolerant and can resist repeated cycles of freezing-thawing [76]. Therefore, the differences in the significance of the olfactory transduction pathways in freeze-tolerance or cryo-resistance observed in boar and giant panda sperm have yet to be clarified.…”

Section: Discussionmentioning

confidence: 99%

Exploration of miRNA and mRNA Profiles in Fresh and Frozen-Thawed Boar Sperm by Transcriptome and Small RNA Sequencing

Dai

Qazi

Ran

et al. 2019

IJMS

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Due to lower farrowing rate and reduced litter size with frozen-thawed semen, over 90% of artificial insemination (AI) is conducted using liquid stored boar semen. Although substantial progress has been made towards optimizing the cryopreservation protocols for boar sperm, the influencing factors and underlying mechanisms related to cryoinjury and freeze tolerance of boar sperm remain largely unknown. In this study, we report the differential expression of mRNAs and miRNAs between fresh and frozen-thawed boar sperm using high-throughput RNA sequencing. Our results showed that 567 mRNAs and 135 miRNAs were differentially expressed (DE) in fresh and frozen-thawed boar sperm. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the majority of DE mRNAs were enriched in environmental information processing such as cytokine-cytokine receptor interactions, PI3K-Akt signaling, cell adhesion, MAPK, and calcium signaling pathways. Moreover, the targets of DE miRNAs were enriched in significant GO terms such as cell process, protein binding, and response to stimuli. In conclusion, we speculate that DE mRNAs and miRNAs are heavily involved in boar sperm response to environment stimuli, apoptosis, and metabolic activities. The differences in expression also reflect the various structural and functional changes in sperm during cryopreservation.

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Section: Discussionmentioning

confidence: 99%

Exploration of miRNA and mRNA Profiles in Fresh and Frozen-Thawed Boar Sperm by Transcriptome and Small RNA Sequencing

Dai

Qazi

Ran

et al. 2019

IJMS

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“…Also, it allows a direct comparison of the shape of these cells among different species. In this way, it is possible to correlate the sperm shape with the parameters of motility, oocyte's ability to attach and penetrate, embryo quality, or even to predict the freezing ratio of a given sperm sample (Gage, 1998;Ros-Santaella, Domínguez-Rebolledo, & Garde, 2014;Santiago-Moreno et al, 2016;Villaverde-Morcillo et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Morphological and morphometric characterization of domestic cat epididymal sperm

Barbosa

Silva

Tabosa

et al. 2019

Reprod Domestic Animals

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Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm2, while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.

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“…Through exploring different freezing diluents (TEST sucrose, egg yolk and glycerol), freezing methods, and thawing solutions, a new procedure for preparing 0.25 mL frozen panda semen was established [ 5 ]. Giant panda sperm appears to be strongly cryo-resistant and can survive repeated cycles of freezing-thawing [ 9 ]. However, the pregnancy rate of insemination using frozen-thawed semen was only 28.57%, which is less than the 33.3% observed in insemination using fresh semen and in natural mating [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

Transcriptome Sequencing Reveals the Differentially Expressed lncRNAs and mRNAs Involved in Cryoinjuries in Frozen-Thawed Giant Panda (Ailuropoda melanoleuca) Sperm

Ran

Zhang

et al. 2018

IJMS

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Sperm cryopreservation and artificial insemination are important methods for giant panda breeding and preservation of extant genetic diversity. Lower conception rates limit the use of artificial insemination with frozen-thawed giant panda sperm, due to the lack of understanding of the cryodamaging or cryoinjuring mechanisms in cryopreservation. Long non-coding RNAs (lncRNAs) are involved in regulating spermatogenesis. However, their roles during cryopreservation remain largely unexplored. Therefore, this study aimed to identify differentially expressed lncRNAs and mRNAs associated with cryodamage or freeze tolerance in frozen-thawed sperm through high throughput sequencing. A total of 61.05 Gb clean reads and 22,774 lncRNA transcripts were obtained. From the sequencing results, 1477 significantly up-regulated and 1,396 significantly down-regulated lncRNA transcripts from fresh and frozen-thawed sperm of giant panda were identified. GO and KEGG showed that the significantly dysregulated lncRNAs and mRNAs were mainly involved in regulating responses to cold stress and apoptosis, such as the integral component of membrane, calcium transport, and various signaling pathways including PI3K-Akt, p53 and cAMP. Our work is the first systematic profiling of lncRNA and mRNA in fresh and frozen-thawed giant panda sperm, and provides valuableinsights into the potential mechanism of cryodamage in sperm.

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Giant panda (Ailuropoda melanoleuca) sperm morphometry and function after repeated freezing and thawing

Cited by 26 publications

References 17 publications

Exploration of miRNA and mRNA Profiles in Fresh and Frozen-Thawed Boar Sperm by Transcriptome and Small RNA Sequencing

Exploration of miRNA and mRNA Profiles in Fresh and Frozen-Thawed Boar Sperm by Transcriptome and Small RNA Sequencing

Morphological and morphometric characterization of domestic cat epididymal sperm

Transcriptome Sequencing Reveals the Differentially Expressed lncRNAs and mRNAs Involved in Cryoinjuries in Frozen-Thawed Giant Panda (Ailuropoda melanoleuca) Sperm

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