RT-qPCR reveals opsin gene upregulation associated with age and sex in guppies (Poecilia reticulata) - a species with color-based sexual selection and 11 visual-opsin genes

Table 1 Guppy housing and rearing

Guppy (n-value) life cycle stage when sequestered in light-dark cycle	Number of offspring in brood	Age at time of sampling^a	Total RNA [ng/μL] ^b and purity (A_260/280) from pooled ^c left eyes	Total RNA [ng/μL] and purity (A_260/280) from pooled right eyes
Primary survey
Gravid female (1)	9 (brood-1)	1-month juveniles ^d	272.9 (2.08)	554.9 (2.09)
Gravid female (1)	12 (brood-2)	1-month juveniles	396.4 (2.10)	429.2 (2.07)
Gravid female (1)	8	2-month juveniles	429.8 (2.10)	470.5 (2.11)
Males (10)	-	Adult	535.0 (2.07)	672.0 (2.10)
Females (6)	-	Adult	441.6 (2.14)	488.3 (2.10)
Secondary survey
Males₁ (5)	-	Adult	544.8 (2.10)
Males₂ (5)	-	Adult	555.6 (2.07)
Males₃ (5)	-	Adult	250.8 (2.09)
Males₄ (5)	-	Adult	463.8 (2.03)
Females₁ (5)	-	Adult	803.4 (2.10)
Females₂ (5)	-	Adult	776.3 (2.10)
Females₃ (5)	-	Adult	158.8 (2.11)
Females₄ (5)	-	Adult	753.2 (2.12)

^a Euthanasia, followed by immediate eye excision, RNA extraction, and cDNA synthesis.
^b Concentration and purity are mean of duplicate measurements within 5ng/μl of each other.
^c Each RNA pool corresponds to 1 of the 18 cDNA samples subsequently generated.
^d 1 month = 28 days.
Note both 1- and 2-month-old juveniles are generally sexually indistinguishable (i.e., not sexually dimorphic); for brevity, the designations 'male' and 'female' are only assigned to adults in this study. Males and females were exposed to each other (i.e., kept in the same aquaria). Male and female guppy numbers (subscripts 1-4) identify different groups.

ISSN: 2730-7182