Physiology and miRNA expression in confined sows with different pupillary light reflex

Figures & data

Table 1. Nutritional standards of formula feed for pregnant sows (%).

CP	CF	CA	Ca	TP	NaCl	Lys
≥15.0%	≤10.0%	≤10.0%	0.5–1.3	≥0.4	0.3–1.0	≥0.5

Diet-nutrient concentration was calculated from the nutrient composition of raw materials in the NRC (2012) (National Research Council, 2012). CP: crude protein; CF: crude fat; CA: crude ash; Ca: calcium; TP: total phosphorus; NaCl: sodium chloride; Lys: lysine.

Table 2. Definitions of pupillary light reflex indicators.

Indicator	Definition
Maximum pupil diameter	The diameter of the pupil before light stimulation
Minimum pupil diameter	The smallest pupil diameter when light stimulated
Contraction rate of pupil	(Maximum pupil diameter − Minimum pupil diameter)/Maximum pupil diameter
Latency of pupillary response	The response time of the pupil’s reaction to light stimulation
Average contraction velocity	The average contraction velocity of the pupil after light stimulation
Maximum contraction velocity	The maximum contraction velocity of the pupil after light stimulation
Average dilation velocity	The average velocity of the pupil from minimum pupil diameter to initial diameter
Time for 75% recovery of the initial pupil diameter	The duration of recovery from minimum pupil diameter to 75% of the initial diameter

Table 3. Primer sequence.

Primer name	Sequence	Size
miR-335-Pig-F	CGCGTCAAGAGCAATAACGAAAAATG	26
miR-1202-Human-F	GTGCCAGCTGCAGTGG	16
miR-16-Pig-F	ATAGCAGCACGTAAATATTGGCG	23
miR-134-Human-F	TGTGACTGGTTGACCAGAGG	20
miR-132-Pig-F	TAACAGTCTACAGCCATGGTCG	22
miR-504-Human-5P	CAGACCCTGGTCTGCACTCTATC	23
miR-504-Human-3P	TATATAGGGAGTGCAGGGCAGGG	23
miR-30e-Human	CGGCTTTCAGTCGGATGTTTACAGC	25
miR-30e-Human-5P	CGCGTGTAAACATCCTTGACTGGAAG	26
miR-34a-Human-5P	CTGGCAGTGTCTTAGCTGGTTGT	23
miR-34a-Human-3P	CGCAATCAGCAAGTATACTGCCCT	24

Table 4. PLR indicators in left and right eyes of sows.

PLR indicators	Left eye	Right eye	p-value
Maximum pupil diameter (mm)	9.82 ± 0.036	9.78 ± 0.038	.44
Minimum pupil diameter (mm)	8.83 ± 0.056	8.85 ± 0.061	.81
Contraction rate of pupil (%)	−0.1 ± 0.004	−0.1 ± 0.005	.47
Latency of pupillary response (s)	0.54 ± 0.018	0.58 ± 0.024	.12
Average contraction velocity (mm/s)	−1.41 ± 0.068	−1.44 ± 0.068	.76
Maximum contraction velocity (mm/s)	−3.22 ± 0.111	−3.05 ± 0.104	.27
Average dilation velocity (mm/s)	0.41 ± 0.03	0.53 ± 0.045	.06
Time for 75% recovery of the initial pupil diameter (s)	1.51 ± 0.153	1.46 ± 0.132	.81

Pupil contraction is opposite to the direction of dilation, “−” indicate the direction of contraction.

Figure 1. Neurophysiological Indicators levels in confined sows with different parity and different pupillary light reflex (PLR) characteristics, including neurophysiological indicators Among Confined Sows with Different PLR (A₁, B₁, C₁), neurophysiological indicators Among Confined Sows with Different Parities (A₂, B₂, C₂). The parities include parity 0 (P0), parity 2 (P2), and parity 5 (P5). The PLR characteristics include weak reflex (WR) and strong reflex (SR). Differences in the (A₁, A₂) serum cortisol (COR) level; (B₁, B₂) serum 5-hydroxytryptamine (5-HT) level; (C₁, C₂) serum dopamine (DA) level among different parity and different PLR groups in confined sows. The standard deviation is expressed by error bars, *p < .05.

Figure 2. Immunoregulatory Factors levels in confined sows with different parity and different pupillary light reflex (PLR) characteristics, including immunoregulatory Factors Among Confined Sows with Different PLR (A₁, B₁, C₁), immunoregulatory Factors Among Confined Sows with Different Parities (A₂, B₂, C₂). The parities include parity 0 (P0), parity 2 (P2), and parity 5 (P5). The PLR characteristics include weak reflex (WR) and strong reflex (SR). (A₁, A₂) serum immunoglobulin A (IgA) level; (B₁, B₂) serum interleukin-6 (IL-6) level; (C₁, C₂) serum tumour necrosis factor-α (TNF-α) level among different parity and different PLR groups in confined sows. The standard deviation is expressed by error bars, *p < .05.

Figure 3. β-EP indicator levels in confined sows with different parity and different pupillary light reflex (PLR) characteristics, including β-EP Among Confined Sows with Different PLR (A₁), β-EP Among Confined Sows with Different Parities (A₂). The parities include parity 0 (P0), parity 2 (P2), and parity 5 (P5). The PLR characteristics include weak reflex (WR) and strong reflex (SR). Differences in the (A₁, A₂) serum beta-endorphin (β-EP) level among different parity and different PLR groups in confined sows. The standard deviation is expressed by error bars, *p < .05.

Figure 4. Relative miRNA expression in sows with different PLR characteristics but the same parity. (A) The relative expression of miR-335; (B) miR-16; (C) miR-134; (D) miR-1202; (E) miR-132; (F) miR-504; (G) miR-30e; and (H) miR-34a. The standard deviation is expressed by error bars, *p < .05; **p < .01.

Figure 5. Relative miRNA expression among sows with different parities but the same PLR characteristics. (A) The relative expression of miR-335; (B) miR-16; (C) miR-134; (D) miR-1202; (E) miR-132; (F) miR-504; (G) miR-30e; and (H) miR-34a. The standard deviation is expressed by error bars, *p < .05.

National Research Council. 2012. Nutrient requirements of swine. 11th ed. Washington DC (WDC): USA National Academies Press.

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