Time course of flinching responses in PND13 rats (males and females combined) in response to an injection of 0.6%, 0.7%, and 0.8% formalin into the plantar surface of the right hindpaw.

<p>Data are presented as mean +/− SE. * <i>p</i><.05; **<i>p</i><.01, *** <i>p</i><.001, against other groups at the same time point.</p

Time course of flinching responses in PND82 rats (males and females combined) in response to an injection of 1.25%, 1.75%, and 2.25% formalin into the plantar surface of the right hindpaw.

<p>Data are presented as mean +/− SE. * <i>p</i><.05, against other groups at the same time point.</p

Time course of licking responses in PND7 rats (males and females combined) in response to an injection of 0.3%, 0.4%, and 0.5% formalin into plantar surface of the left hindpaw.

<p>Data are presented as mean +/− SE. * <i>p</i><.05; **<i>p</i><.01; *** <i>p</i><.001, against other groups at the same time point.</p

Time course of licking responses in PND82 male (A) and female (B) rats in response to an injection of 1.25%, 1.75%, and 2.25% formalin into the plantar surface of the right hindpaw.

<p>Baseline is the time prior to formalin injection (10 min). Data are presented as mean +/− SE. * <i>p</i><.05, **<i>p</i><.01, against other groups at the same time point.</p

Time course of licking responses in PND22 male (A) and female (B) rats in response to an injection of 0.9%, 1%, and 1.1% formalin into the plantar surface of the left hindpaw.

<p>Data are presented as mean +/− SE. * <i>p</i><.05; **<i>p</i><.01, against other groups at the same time point.</p

Summary of GR isoform expression in guinea pig placenta in relation to fetal sex, gestational age and betamethasone exposure.

<p>The circles represent a cell with the smaller circle being the nucleus and the larger being the cytoplasm. Isoforms that were present under each condition are in small font while isoforms that were significantly altered by treatment, or gestational age are in bold. Fig 4a represents female placentae and 4b represents male placentae. This figure was generated from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148226#pone.0148226.t002" target="_blank">Table 2</a> and isoforms were included in the figure if the median value was greater than 0 regardless of whether GR expression was significantly altered or not.</p

Maternal and Neonatal Characteristics.

<p>Maternal and Neonatal Characteristics.</p

Identification of Eight Different Isoforms of the Glucocorticoid Receptor in Guinea Pig Placenta: Relationship to Preterm Delivery, Sex and Betamethasone Exposure

<div><p>The placental glucocorticoid receptor (GR) is central to glucocorticoid signalling and for mediating steroid effects on pathways associated with fetal growth and lung maturation but the GR has not been examined in the guinea pig placenta even though this animal is regularly used as a model of preterm birth and excess glucocorticoid exposure. Guinea pig dams received subcutaneous injections of either vehicle or betamethasone at 24 and 12 hours prior to preterm or term caesarean-section delivery. At delivery pup and organ weights were recorded. Placentae were dissected, weighed and analysed using Western blot to examine GR isoform expression in nuclear and cytoplasmic extracts. A comparative examination of the guinea pig GR gene identified it is capable of producing seven of the eight translational GR isoforms which include GRα-A, C1, C2, C3, D1, D2, and D3. GRα-B is not produced in the Guinea Pig. Total GR antibody identified 10 specific bands from term (n = 29) and preterm pregnancies (n = 27). Known isoforms included GRγ, GRα A, GRβ, GRP, GRA and GRα D1-3. There were sex and gestational age differences in placental GR isoform expression. Placental GRα A was detected in the cytoplasm of all groups but was significantly increased in the cytoplasm and nucleus of preterm males and females exposed to betamethasone and untreated term males (KW-ANOVA, P = 0.0001, P = 0.001). Cytoplasmic expression of GRβ was increased in female preterm placentae and preterm and term male placentae exposed to betamethasone (P = 0.01). Nuclear expression of GRβ was increased in all placentae exposed to betamethasone (P = 0.0001). GRα D2 and GRα D3 were increased in male preterm placentae when exposed to betamethasone (P = 0.01, P = 0.02). The current data suggests the sex-specific placental response to maternal betamethasone may be dependent on the expression of a combination of GR isoforms.</p></div

A preabsorption control was conducted to identify nonspecific binding of the antibody.

<p>GR antibody was preabsorbed with control peptide prior to Western Blot exposure. The left panel represents binding after GR antibody preabsorption and the right panel is the blot exposed to GR antibody alone. Human placental tissues was included as a positive control.</p

Mental health status at commencement and completion of the trial for App-users.

<p>Mental health status at commencement and completion of the trial for App-users.</p