INTRODUCTION:

Locomotor activity is spontaneous movement of animals from one place to another place. The ability of an organism to move from one place to another through coordinated limb movement is referred to as locomotion. Locomotion is a fundamental skill required by animals and humans for a large variety of actions and it’s a complex neural process characterized by rhythmic activity using varied degrees of freedom. Locomotion control is organized in a way that the neural network in the spinal cord generates basic rhythmic patterns necessary for movement, while the higher centres of the brain interact with the spinal circuits for posture control and accurate limb movements.

The locomotor behaviour of zebrafish has been widely used to study neuro-behaviour^1,2,3.

In between top 10 deadly diseases CNS depression is one of them. CNS depression can be treated by proper counseling and medication but total cure is nearly impossible still today. It was estimated that about 200 million people worldwide suffered from CNS depression in 2010, and it is expected to reach 300 million by 2025. People in low-to middle-income countries are more likely suffer from CNS depression due to high price of allopathic drug. As India is a middle-income country that why our country will much affected in coming days if a better step can’t taken against CNS depression.

People with CNS depression often use herbal medicine for treatment. Many people say these treatments help them to cope better with CNS depression. Research is looking into whether some herbs or plant treatments used in herbal medicine could help to prevent or treat CNS depression.

The zebrafish (Danio rerio) is a popular model organism for behavioural genetics and developmental studies, especially forward genetics because the nervous system of the zebrafish exhibits developmentally, structurally, functionally and pharmacologically conserved similarities with the mammalian nervous system, zebrafish are also widely used as experimental animals in neural and neuropharmacological studies. As early as 4-5 days post fertilization (dpf), zebrafish larvae develop different behaviours, including emotions (anxiety and fear), learning and memory, goal-directed behaviour, optokinetic responses, and pre-pulse inhibition. Neuroactive drugs that interact with different receptors of the central nervous system result in diverse behavioural manifestations in zebrafish drugs inhibited their locomotor activities for zebrafish, changes in illumination conditions usually result in changes in the levels of locomotor activities⁴.

It has been shown that the locomotor activities of larval zebrafish were significantly enhanced when the illumination changed from light to dark, and some neuroactive drugs were able to modify or even reverse this phenomenon. A visual motor response (VMR) assay that focused on the Instantaneous changes in the swimming activities of zebrafish in response to changes in lighting conditions was previously described. To characterize drug effects on locomotor activities, instead of focusing on the transient instantaneous locomotor changes caused by changes in the lighting conditions, we aimed to examine the more sustained changes in the swimming activities that are caused by differences in lighting, i.e., light and dark conditions. However, studies thus far have primarily focused on the qualitative aspect of this phenomenon; therefore, a new parameter-the lighting motor index-was devised in this work to focus on the quantitative aspect of this phenomenon. In this study, we examined the influences of different concentrations of various neuroactive drugs on the locomotor activities and the lighting motor Indexes of the zebrafish larvae under light and dark conditions. With this comparative study, we attempted to speculate a correlation between the drug activities and their behavioural influences to promote the identification of the activities of novel drugs through their effects on zebrafish locomotion^5,6,7.

The locomotor activity of methanolic extract of Erigeron bonariensis can be help to establish a new drug against CNS illness which is not reported till today. Hence, aim of the present study is to designed for locomotor activity of methanolic extract of Erigeron bonariensis on zebra fish model⁸.

MATERIALS AND METHODS:

Collection of plant:

The plant of Erigeron bonariensis was collected from a field in Haldia, West Bengal and also the plant was identified as the same by the scientist incharge, Central national herbarium, Botanical survey of India, Kolkata, West Bengal.

Extraction of plant sample:

The plants were thoroughly washed and dried under shade for 10 days^9,10. The various plant parts were separated and grounded into powder using and electrically operated grinder. Powdered plant material (100gm) was mixed with 200ml of Methanol and kept in an Iodine flask for 72 hours with often everyday stirring it with a glass rod. The extract was filtered with Whatmann’s No 1 paper; filtrate was evaporated to dryness in sunlight¹¹.

Phytochemical screening of plant sample:

Preliminary phytochemical study of alkaloids, carbohydrates, glycosides, protein, saponin, phenolic compound and tannins were performed^10,12,13.

Preparation of test sample:

Three sample of different concentration (1 mg/ml, 2 mg/ml, 4 mg/ml) was prepared through serial dilution¹⁴.

Zebrafish care:

All zebrafish were acclimated to constant laboratory conditions (14 h light:10 h dark photoperiod, diet, water, 28 C) for at least one week in stock aquaria before experiments were conducted. Adult zebrafish were maintained in tap water conditioned chlorine. Fish were fed twice daily withfish flakes (0.5 mg/fish/day)^17,18,19.

Grouping of animals:

Experimental zebrafish weredivided into 4 Groups of 6 animals each (n=6). Group 1 – served as control, Group 2 Group 3 and Group 4 – were treated with test substance at three dose (1 mg/ml, 2 mg/ml, 4 mg/ml) levels. divided into three groups control group and test group (Different groups for different concentration)^14,15,16.

Pharmacological studies:

Locomotor activity (Total distance travelled and speed for 1minute is record) was studied. For this at first the tank was filled with purified water. Then24 Zebra fishes were selected (6 for control group & 6 for3 types of each test group). Next One by one control group fishes were put in the tank for swimming without the extract and the total distance travelled and speed for 1minute was recorded/ measured. Then 1mg/ml sample was put in the water drop by drop (4-5 drops) & left for few minutes. Then the total distance travelled & speed was observed and calculated for 1 minute under the influence of 1mg/ml sample for the first test group. The above process was carried out for the next two test group fishes for the concentration (2mg/ml, 4mg/ml) for 1 minute. Then the possible increase/decrease in the distance travelled and speed was observed and the graph was plotted for the same^20,21,22.

RESULTS:

Table 1: Phytochemical screening of plant

Sl No.	Phytochemical test	Observation
1	Alkaloids	-
2	Carbohydrate	-
3	Glycosides	+
4	Flavonoids	+
5	Tannins	+
6	Steroids & Terpenoids	+
7	Amino acid	+
8	Saponin	-

Table 2: Behavioral pattern by total distance travelled

Sl No.	Concentration (mg/ml)	Control Group	Test Group
Sl No.	Concentration (mg/ml)	Distance (m)	Distance (m)
1	1	4.4	6.16
2	2	4.8	7.04
3	4	4.4	7.92

Figure 1: Total distance travelled control vs test

Table 3: Behavioral pattern by average speed

Sl No.	Concentration (mg/ml)	Control Group	Test Group
Sl No.	Concentration (mg/ml)	Speed (m)	Speed (m)
1	1	0.07	0.102
2	2	0.08	0.117
3	4	0.07	0.132

Figure 2. Average speed control vs test

DISCUSSION:

The study showed the presence of glycosides, flavonoids, Amino acid, tannins and absence of Alkaloids, carbohydrates and saponin. In this study the total distance by Zebra fish was calculated for 1 minute. The total distance travelled by control group and the 1st test group (for 1mg per ml) was 4.4 m and 6.16 m respectively. The total distance travelledby control group and the 2nd test group (for 2mg per ml) was 4.8 m and 7.04 m respectively. The total distance travelled by control group and the 3^rdtest group (for 4mg per ml) was 4.4 m and 7.92 m respectively. The distance travelled has increased in the test group as compare to the control group, since speed is the ratio of distance travelled to time taken hence the speed will also increase. The control and the test group (for 1mg per ml) travelled at the speed of 0.07 m/sec and 0.102 m/sec respectively. The control and the test group (For 2mg per ml) travelled at the speed of 0.08 m/sec and 0.117 m/sec respectively. The control and the test group (For 4mg per ml) travelled at the speed of 0.07 m/sec and 0.132 m/sec respectively.

The present research study can be concluded that, after administration of the methanolic extract in water with different concentration (1mg/ml, 2mg/ml, 4mg/ml) to the Zebra fish, the total distancetravelled by the test group is higher than the control group. The speed attained by the test group is higher than the control group. The experiment also observed that with the increase in the concentration of the sample, the total distance travelled and Speed also increased. So, it is concluded that the locomotor activity occurs. Hence, the plant has CNS Stimulant activity may be due presence of different phytochemical constituents.

CONFLICT OF INTEREST:

NIL

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Received on 14.09.2022 Modified on 11.02.2023

Res. J. Pharmacology and Pharmacodynamics. 2023;15(2):45-48.

DOI: 10.52711/2321-5836.2023.00009