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41.
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47.
  • Aanesen, Arthur (författare)
  • GABA and human spermatozoa : characterization and regulation of GABA transport proteins
  • 1998
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • The present project aimed at investigating the interaction between GABA and human spermatozoa under in vitro conditions. Our initial hypothesis was that human spermatozoa had specific binding proteins for GABA and that the binding of GABA to such binding sites could affect sperm fimction. Studies on swim-up preparations of human spermatozoa incubated with radiolabelled GABA in the presence of unlabelled GABA, alternatively displacers of GABAA/B receptors and GABA transport proteins. indicated that GABA specific binding sites were present on the surface of human spermatozoa. The potency of the different inhibitors indicated that the binding sites possibly represented GABA transport proteins. No effect on sperm motility or AR was observed following incubation with GABA. Further studies indicated that human spermatozoa are capable of a carrier mediated GABA uptake. The uptake is dependent on the concentration of chloride and sodium in the external medium, and the kinetic properties of the carrier resembled high affinity GABA transport proteins. Uptake of radiolabelled GABA into human spermatozoa in the presence of steroids was investigated. We examined progesterone, other steroids known to increase calcium influx, and steroids known to be ineffective as stimulators of calcium influx in human sperm. The results demonstrated a twofold increase in GABA uptake following preincubation with P or steroids known to stimulate calcium influx. In contrast, an inhibition of GABA uptake following preincubation with activators of protein kinase C was observed. The results suggest a role for PKC in the regulation of GABA transport in human spermatozoa. The reduced GABA uptake following incubation with phorbol esters and diacyl glycerol (DAG) analogues, as well as the lack of effect observed following addition of inactive DAG analogues, strongly indicate such a regulatory pathway. The existence of GABA transporter m-RNA and proteins in human testis and sperm was examined using RT-PCR, immunohistochemistry and immunoblotting. Oligonucleotide primer pairs were designed from nucleotide sequences for the three cloned GABA transport proteins identified in human brain: GAT-1, GAT-3 and BGT-1. From homogenized human testis, PCR products of predicted size and with homology to respective GABA transport proteins were identified. This finding indicates the MRNA expression of all three GABA transporters in human testicular tissue. Studies on the protein expression of GABA transporters in sperm cells were performed with SDS-PAGE and immunoblotting techniques. Immunoblots using polyclonal antibodies raised against rat GAT-1, GAT-2 and GAT-3 recognized bands of approximately 65kDa, indicating cross-reactivity to GABA transporters present in human sperm cells. Pharmacological characterization of GABA uptake with specific inhibitors of GABA transport revealed a unique sequence in the relative potencies of the inhibitors tested. The results may indicate the presence of more than one GABA transporter in the plasma membrane of human spermatozoa. Immunoblots and immunohistochemistry studies indicate the presence of one or several GABA transporters in human spermatozoa and testis, these results are supported by pharmacological studies using specific inhibitors of GABA transport. These studies add new information to the present knowledge concerning GABA and human spermatozoa. They also for the first time describe specific GABA transport proteins in the male reproductive tract. Further studies are however, required to elucidate the functional role for these proteins in terms of human reproduction.
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50.
  • Aanesen, Arthur, et al. (författare)
  • Prospective study of a Swedish infertile cohort 2005-08 : population characteristics, treatments and pregnancy rates
  • 2014
  • Ingår i: Family Practice. - 0263-2136. ; 31:3, s. 290-7
  • Tidskriftsartikel (refereegranskat)abstract
    • BACKGROUND: We here report on results from a prospective study comprising 380 infertile couples undergoing infertility work-up and various treatments for infertility in our clinic. The aim was to investigate the overall birth rate as a result of different treatments, as well as spontaneous pregnancies.METHODS: Three hundred and eighty couples were consecutively included between December 2005 and May 2008. All couples underwent a fertility work-up, including hysterosalpingogram, hormonal characterization, clinical examination, screening for infectious diseases and semen analysis. The mean age of the women at the time of inclusion was 33.2 years. The mean duration of infertility prior to inclusion was 1.8 years. And 46.6% (n = 177) of the women had been pregnant prior to their first visit to the clinic and 30.0% (n = 114) had been pregnant earlier in their present relationship.RESULTS: As of November 2010, 57.3% (n = 218) of the women had given birth to a child when they were lost to follow up by the study. Spontaneous conception was observed in 11.3% (n = 43) of the women, 14.5% (n = 64) conceived after intrauterine insemination (IUI), 4.2% (n = 16) conceived after ovarian hyperstimulation and ovulation induction (OH/OI) and 28.4% (n = 113) after in vitro fertilization. There were 280 pregnancies and 58 spontaneous abortions (22.3%) in the group. Mean anti-mullerian hormone significantly correlated with antral follicle count and age and was significantly higher in the subgroup that became pregnant after IUI.CONCLUSIONS: Spontaneous pregnancies and IUI + OH/OI contributed significantly to the pregnancies observed in the total population. Predictive factors for pregnancy were anti-mullerian hormone in the group undergoing IUI treatment and in the age group ≥38-duration of infertility. Previous pregnancies, body mass index, estradiol, follicle stimulating hormone or having given birth prior to the infertility period were not predictive of pregnancy for the infertile couples in this study.
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