1. Brain-derived neurotrophic factor is a regulator of human oocyte maturation and early embryo development 
Richard A. Anderson, M.D., Ph.D., Rosemary A.L. Bayne, Ph.D., John Gardner, M.Sc., Paul A. De Sousa, Ph.D. 
Fertility and Sterility 
Volume 93, Issue 5, Pages (March 2010)
DOI: /j.fertnstert
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Detection of neurotrophin ligand and receptor messenger RNA (mRNA) expression. Representative reverse transcriptase–polymerase chain reaction (RT-PCR) results for analysis of expression of brain-derived neurotrophic factor (BDNF) and its receptors in immature and mature cumulus, and in oocytes. Imm = immature cumulus from cumulus–oocyte complexes (COCs) aspirated from small antral follicles without in vitro maturation. In vivo matured = cumulus from COCs from intracytoplasmic sperm injection (ICSI) cycles (i.e., after superovulation and hCG administration). In vitro matured = cumulus from immature COCs after 24 hours of in vitro maturation in hormones + serum replacement. Receptors analyzed were p75NGFR, full-length TrkB (flTrkB), and the truncated isoform of TrkB (trTrkB). M = molecular weight marker; C = positive control tissue.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Cumulus expansion in in vitro maturation treatment groups. Representative examples of cumulus–oocyte complexes before (A, C) and after (B, D) maturation in vitro for 44 hours, exhibiting cumulus expansion in the latter. Shown are results from maturation in media supplemented with hormones + serum replacement (A, B) and brain-derived neurotrophic factor + serum replacement (C, D). Cumulus expansion varied from a typical halo formation of cells around oocytes (B) to virtual denuding of oocytes owing to self-clustering of expanded cumulus, both of which were observed in all treatment conditions supplemented with serum replacement, but not in its absence. Bar = 100 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Polar body emission and parthenogenetic embryo development after in vitro maturation. Representative examples of polar body emission in oocytes (A, B) after 44 hours in in vitro maturation, or parthenogenetic embryo development after 2 days (C–G) or 5–7 days (H–J). polar body emission was evaluated by fluorescence staining for cortical microfilaments (A), microtubules (A'), and DNA (A”), or phase microscopy (B). Embryo development was evaluated by phase (C–H) or fluorescence (I, J) microscopy, the latter to assess nuclear morphology and number. No differences in polar body or oocyte morphology were apparent across different treatment groups (shown are oocytes matured in hormones + serum replacement (H + SR) (A) and brain-derived neurotrophic factor + serum replacement (BDNF + SR) (B). At day 2 after activation, cleavage was characterized as normal (C, D), which occasionally allowed for a minor degree of cytoplasmic fragmentation (E, middle embryo), abnormal (asterisked embryos in E, F, G), or uncleaved (see E and G). Limited development of compacted morulae and blastocysts was observed by 5–7 days after activation (H, with corresponding Dapi-stained nuclei of subset of embryos in I, J). Shown are parthenogenetic embryos derived from oocytes matured in H + SR (C), H + BDNF (D), BDNF + SR (E), H + BDNF/AB (F), H + SR + TrkB/Fc (G). Bar = 100 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Quantification of polar body (PB) emission and parthenogenetic embryo development in in vitro maturation treatment groups. Effect of in vitro maturation on PB emission and day 2 embryo cleavage after parthenogenetic activation. (A) percentage of PB emission; (B) normal cleavage; (C) abnormal cleavage/fragmentation; (D) uncleaved embryos as percentage of number of cumulus–oocyte complexes (COCs). Horizontal bars denote statistically significant differences between treatment groups. ∗P<.05; ∗∗P<.01. Number of COCs in each treatment group: H + SR, 50; H + BDNF, 45; H + BDNF + SR, 26; BDNF + SR, 15; H + BDNF/AB, 22; H + SR + TrkB/Fc, 7. See Figure 3 legend for other abbreviations.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

6. Figure 5
Assessment of cumulus gene expression during in vitro maturation (IVM). Effect of IVM on expression of HAS2, TNFA1P6, BDNF, PTGS2, and GREM1 in immature cumulus, after IVM and after in vivo maturation. Imm = immature cumulus from cumulus–oocyte complexes on aspiration from small antral follicles. H + SR = cumulus after 24 hours in IVM in medium containing hormones and serum replacement; BDNF + SR = cumulus after 24 hours in IVM in medium containing BDNF and serum replacement; IVF = in vivo-matured cumulus from cumulus–oocyte complexes aspirated after superovulation/hCG administration. See text and Figure 3 legend for other abbreviations. Mean ±SEM, n = 4–6 per group. Gene expression is expressed relative to GAPDH. Horizontal bars denote statistically significant differences between treatment groups, ∗P<.05; ∗∗P<.01. Note: PTGS2 is plotted on a log scale.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions