Advanced search options

Advanced Search Options 🞨

Browse by author name (“Author name starts with…”).

Find ETDs with:

in
/  
in
/  
in
/  
in

Written in Published in Earliest date Latest date

Sorted by

Results per page:

Sorted by: relevance · author · university · dateNew search

You searched for subject:(PLC zeta). Showing records 1 – 2 of 2 total matches.

Search Limiters

Last 2 Years | English Only

No search limiters apply to these results.

▼ Search Limiters


Colorado State University

1. Gonzalez-Castro, Raul A. Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection.

Degree: PhD, Biomedical Sciences, 2018, Colorado State University

When performing intracytoplasmic sperm injection (ICSI), many in vivo mechanisms for sperm selection are bypassed; however, sperm must still be capable of activating the oocyte for successful fertilization. Limited information is available for horses on the effect of sperm preparation method and sperm characteristics that affect ICSI outcome. The overall objectives of this dissertation were to: 1) study the association between sperm sorting methods, sperm population characteristics, and equine ICSI outcome, and 2) characterize sperm oocyte activating factors in stallion sperm, such as phospholipase C zeta (PLCz) and postacrosomal WW binding protein (PAWP). In Experiment 1, a microfluidic device was used to sort frozen-thawed sperm from stallions (n=19), which resulted in a sperm subpopulation with improved motility, morphology, viability and DNA integrity (P<0.05) compared to the original sample. Then, microfluidic sorting was compared with the swim-up procedure and density gradient centrifugation. Swim-up was the least effective method to separate equine sperm. Microfluidic sorting and density gradient centrifugation sorted a sperm subpopulation with similar parameters, improving motility, viability and DNA integrity. After ICSI (n=45), no differences (P>0.3) were observed for cleavage and embryo development among sorting methods. In Experiment 2, sperm population parameters from which individual sperm were selected for injection were analyzed immediately after ICSI and correlated with the outcome. Sperm morphology, viability, membrane integrity measurement of hypoosmotic swelling and DNA integrity were evaluated in frozen-thawed sperm (n=114) used for ICSI in a program. Among sperm parameters, viability correlated positively with normal morphology and membrane integrity (P<0.05). Normal sperm morphology and DNA integrity were not predictive of ICSI outcome. Viability was predictive of cleavage and blastocyst formation, and membrane integrity was predictive of early pregnancy (P<0.05). In Experiment 3, PLCz and PAWP were identified, localized and quantified in stallion sperm, and the relationship with other sperm parameters was investigated. PLCz was identified as a 71 kDa protein and located in the acrosomal and postacrosomal region, midpiece and principal piece of the tail. PAWP was identified by two bands of ~28 and ~32 kDa, located in the postacrosomal region, midpiece and principal piece of the tail. The expression of PLCz and PAWP correlated positively (P=0.04) when analyzed for sperm of 14 stallions. Flow cytometric assessment was feasible for PLCz, but not for PAWP. Expression and percentages of positive labeled sperm for PLCz varied among stallions (n=21). Expression of PLCz was higher in live than dead sperm (P<0.005), and DNA fragmentation correlated negatively with PLCz expression (P<0.04). In conclusion, microfluidic sorting and density gradient centrifugation resulted in a subpopulation of sperm with high quality parameters for ICSI. The probability of sperm-injected oocytes to develop into an embryo… Advisors/Committee Members: Carnevale, Elaine (advisor), Grahman, James (advisor), Seidel, George (committee member), Gerrit, Bouma (committee member), Ann, Hess (committee member).

Subjects/Keywords: microfluidics; PLC zeta; stallion; PAWP; ICSI; sperm

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Gonzalez-Castro, R. A. (2018). Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/191446

Chicago Manual of Style (16th Edition):

Gonzalez-Castro, Raul A. “Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection.” 2018. Doctoral Dissertation, Colorado State University. Accessed April 16, 2021. http://hdl.handle.net/10217/191446.

MLA Handbook (7th Edition):

Gonzalez-Castro, Raul A. “Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection.” 2018. Web. 16 Apr 2021.

Vancouver:

Gonzalez-Castro RA. Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection. [Internet] [Doctoral dissertation]. Colorado State University; 2018. [cited 2021 Apr 16]. Available from: http://hdl.handle.net/10217/191446.

Council of Science Editors:

Gonzalez-Castro RA. Characterization of equine sperm attributes and selection for intracytoplasmic sperm injection. [Doctoral Dissertation]. Colorado State University; 2018. Available from: http://hdl.handle.net/10217/191446


University of Oxford

2. Amdani, Siti Nornadhirah. The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency.

Degree: PhD, 2018, University of Oxford

Oocyte activation deficiency (OAD) is an infertile condition observed in patients who have experienced recurrent total fertilisation failure (TFF) following intracytoplasmic sperm injection treatment. This condition was considered to be an idiopathic factor for a long time but strong clinical evidence now suggests that dysfunctional forms of phospholipase C zeta (PLCζ) may be predominant causative factors for OAD. Genetic contribution has played a role in patients suspected of having OAD, as four PLCζ exonic mutations have been discovered and characterised as being the cause of infertility. In this study, a novel nonsense mutation, PLCζK322Stop, was identified in the PLCζ XY-linker region of Patient LR. This variant results in the truncation of approximately half of PLCζ, therefore was non-functional when activity was tested. Patient LR, which also exhibited a previously reported mutation, PLCζH233L, may suggest that the patient is sub-fertile, as opposed to being infertile, as initially expected. Although research has purely focused upon the coding regions of PLCζ, it was obvious that our knowledge of PLCζ regulatory elements remain very limited. Next generation sequencing (NGS) was therefore employed to detect variants in the non-coding regions of PLCζ, promoter and introns, which may have resulted in the observed phenotypic diversity of PLCζ expression in fertile and infertile patients. As a result of mapping failure, an alternative approach was considered to identify variants within human PLCζ, and this involved using the single nucleotide polymorphism (SNP) database. Over 2500 SNPs were localised in the intronic regions of PLCζ and thus, it could be speculated that these variants may help elucidate the wide variation of PLCζ expression reported. Additionally, two particular patients with TFF (79 and 107) were investigated in this study to identify an association with PLCζ and their infertile state. For Patient 79, multiple PLCζ immunofluorescence analysis was performed and a significant improvement in PLCζ expression was observed one year after his first investigation. This may have been the result of an external factor, which influenced protein expression. As for Patient 107, a novel substitution mutation, PLCζV193E, was identified and was predicted to affect PLCζ stability and folding. There is global interest to create a safer and alternative OAD therapy, namely a human recombinant PLCζ protein (hrPLCζ). The first method, using a bacterial cell line resulted in successful purification and identification but the product proved to be inactive following mouse oocyte microinjection. The second method involved production of a mammalian-expressed hrPLCζ, which was successfully purified and identified but due to time restrictions, could not be tested for functionality. Concurrently, the findings in this thesis have reinforced the association between PLCζ and OAD, and provided improved options for the…

Subjects/Keywords: 612.6; next generation sequencing (NGS); variants; promoter; phospholipase C zeta (PLC?); human recombinant PLC? protein (hrPLC?); introns; Oocyte activation deficiency (OAD)

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Amdani, S. N. (2018). The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency. (Doctoral Dissertation). University of Oxford. Retrieved from http://ora.ox.ac.uk/objects/uuid:af4c4f98-497a-4666-9eec-a46bb579dd59 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.736166

Chicago Manual of Style (16th Edition):

Amdani, Siti Nornadhirah. “The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency.” 2018. Doctoral Dissertation, University of Oxford. Accessed April 16, 2021. http://ora.ox.ac.uk/objects/uuid:af4c4f98-497a-4666-9eec-a46bb579dd59 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.736166.

MLA Handbook (7th Edition):

Amdani, Siti Nornadhirah. “The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency.” 2018. Web. 16 Apr 2021.

Vancouver:

Amdani SN. The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency. [Internet] [Doctoral dissertation]. University of Oxford; 2018. [cited 2021 Apr 16]. Available from: http://ora.ox.ac.uk/objects/uuid:af4c4f98-497a-4666-9eec-a46bb579dd59 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.736166.

Council of Science Editors:

Amdani SN. The oocyte-activation factor, phospholipase C zeta (PLCζ) : clinical prognosis, diagnosis, and treatment of oocyte activation deficiency. [Doctoral Dissertation]. University of Oxford; 2018. Available from: http://ora.ox.ac.uk/objects/uuid:af4c4f98-497a-4666-9eec-a46bb579dd59 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.736166

.